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By Ekasit Nisayan

A Thesis Submitted in Partial Fulfillment of the Requirements for the Degree MASTER OF SCIENCE Department of Biology Graduate School SILPAKORN UNIVERSITY 2010 ´–”·˜ª·š¥µ¨´¥ ¤®µª·š¥µ¨´¥«·¨žµ„¦ °œ»¤´˜·Ä®oª·š¥µœ·¡œ›rÁ¦ºÉ°Š “„µ¦Á¡µ³Á¨¸Ê¥Š ÁœºÊ°Á¥ºÉ°„¨oª¥Å¤o¦°ŠÁšoµœµ¦¸ (Paphiopedilum ‘Delrosi’)” Á­œ°Ã—¥ œµ¥Á°„­·š›·Í œ·­¥´œ˜r Áž}œ ­nªœ®œ¹ÉŠ °Š„µ¦«¹„¬µ˜µ¤®¨´„­¼˜¦ž¦·µª·š¥µ«µ­˜¦¤®µ´–”·˜ ­µ µª·µ¸ªªš¥µ·

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­nŠÁ­¦·¤Ä®oÁ„·—¥°—­¼Š 1.6 ¥°—˜n°¥°—Á¦·É¤˜oœ ¨³Ä®oÁž°¦rÁŽÈœ˜r„µ¦Á„·—¥°— 90 Áž°¦rÁŽÈœ˜r œ°„‹µ„œ¸Ê „µ¦´„œÎµ„µ¦Á„·—¥°—‹µ„­nªœ o° ¡ªnµœÊ嘵¨Ž¼Ã‡¦­ 20 „¦´¤˜n°¨·˜¦ š¸Éž¦µ«‹µ„­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ ¤¸‡ªµ¤Á®¤µ³­¤Äœ„µ¦´„œÎµ„µ¦Á„·—¥°—‹µ„­nªœ o° Ž¹ÉŠÄ®oÁž°¦rÁŽÈœ˜r„µ¦Á„·—¥°— 100 Áž°¦rÁŽÈœ˜r ­Îµ®¦´„µ¦Á¡µ³Á¨¸Ê¥Š·Êœ­nªœÁœºÊ°Á¥ºÉ°¦³®ªnµŠ o° ¨³·Êœ­nªœÄ œ­¼˜¦°µ®µ¦ 1/2 MS (1/2 strength macro-and full-strength micro elements) š¸Éž¦³„°—oª¥œÊ嘵¨Ž¼Ã‡¦­ „¨¼Ã‡­ ¨³¤°¨Ã˜­ 20 „¦´¤˜n° ¨·˜¦ ¦nª¤„´ TDZ (0.45-4.54 ŤÇ¦Ã¤¨µ¦)r Á¡¸¥Š°¥nµŠÁ—¥ª¸ ®¦°¦º ª¤„n ´ 2,4-D (4.52-13.56 ŤÇ¦Ã¤ ¨µ¦)r ¡ªnµÅ¤n¤¸„µ¦´„œÎµÄ®oÁ„·—‡¨¨´­Â¨³ somatic embryos š´ÊŠ‹µ„·Êœ­nªœ¦³®ªnµŠ o° ®¦°º ·Êœ­nªœÄ š¸ÉÁ¡µ³Á¨¸Ê¥Šœ°µ®µ¦š»„­¼˜¦ ®¨´Š‹µ„„µ¦Á¡µ³Á¨¸Ê¥ŠÁž}œÁª¨µ 4-5 ­´ž—µ®r ÁœºÊ°Á¥ºÉ°š¸ÉÁ¡µ³Á¨¸Ê¥ŠÁž¨¸É¥œÁž}œ­¸ œÊ嘵¨Â¨³˜µ¥Äœš¸É­»—

£µ‡ª·µ¸ªª·š¥µ ´–”·˜ª·š¥µ¨´¥ ¤®µªš¥µ¨· ´¥«·¨žµ„¦ že„µ¦«¹„¬µ 2553 ¨µ¥¤º°ºÉ°œ´„«„¬µ¹ ...... ¨µ¥¤º°ºÉ°°µ‹µ¦¥rš¸Éž¦¹„¬µª·š¥µœ·¡œ›r 1...... 2...... Š 50303208 : MAJOR : BIOLOGY KEY WORD : LADY'S SLIPPER ORCHID/PROPAGATION/TISSUE CULTURE EKASIT NISAYAN : TISSUE CULTURE OF THE LADY’S SLIPPER ORCHID (PAPHIOPEDILUM ‘DELROSI’). THESIS ADVISORS : ASST.PROF.CHOCKPISIT THEPSITHAR, Ph.D.,AND ASSOC.PROF. AREE THONGPUKDEE,Ph.D. 94 pp.

Micropropagation of Lady’s slipper orchid (Paphiopedilum ‘Delrosi’) was established. Multiple shoots were induced from a shoot, a stem without shoot and a node while calli or somatic embryos were induced from an internode and a leaf segment on different types and concentrations of growth regulator in culture media for 16 weeks. An individual shoot was cultured on modified Hyponex media [3 g/l Hyponex (6.5N-4.5P-19K), 2 g/l peptone, 100 g/l boiled potato juice and 20 g/l sucrose] supplemented with thidiazuron (TDZ), 6-benzyladenine (BA), or kinetin (Kn) or combination with 2,4-dichlorophenoxyacetic acid (2,4-D) and adenine sulfate (Ads) at various concentrations. It was found that 4.65 μM Kn in combination with 4.52 μM 2,4-D and 0.45 μM TDZ in combination with 4,52 μM 2,4-D provided high percentages of shooting efficiency and number of new shoots (100% with 1.4 new shoots per explant and 90% with 1.2-1.3 new shoots per explant, respectively). Studying on effects of various sugars on multiple shoot induction formed that 20 g/l glucose in combinations with 0.45 μM TDZ and 4.52 μM 2,4-D resulted in the highest new shoot numbers, 1.8 new shoots per explant, with 80% shooting efficiency. For multiple shoot induction from a stem without shoot, 20 g/l sucrose in combinations with 4.65 μM Kn and 4.52 μM 2,4-D promoted high number of shoots, 1.6 new shoots per explant, with 90% shooting efficiency. Furthermore, 20g/l sucrose in medium without plant growth regulator was suitable for shoot induction from a node with 100% shooting efficiency. For in vitro culture of an internode and leaf segment cultured for 16 weeks on modified Murashige and Skoog (MS) medium supplemented with TDZ (0.45-4.54 μM) singly or in combination with 2,4-D (4.52- 13.56 μM). It was shown that there was no callus and somatic embryos induced from either an internode or a leaf segments culture on all media. After 4-5 weeks of culturing, explants turned brown and eventually died.

Department of Biology Graduate School, Silpakorn University Academic Year 2010 Student's signature ...... Thesis Advisors' signature 1...... 2...... ‹ „·˜˜·„¦¦¤ž¦³„µ«

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«¹„¬µŸ¨ °Š­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ TDZ BA Kn ¨³ 2,4-D ˜n°„µ¦´„œÎµ„µ¦Á„·—¥°—šª¸‡¼–‹µ„­ªœ¥°— °Š„¨n oª¥Å¤o Paphiopedilum ‘Delrosi’...... 29 «¹„¬µœ·— °ŠœÊ嘵¨š¸ÉÁ®¤µ³­¤˜n°„µ¦´„œÎµ„µ¦Á„—¥°—šª· ¸‡¼–‹µ„

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«¹„¬µ­¼˜¦°µ®µ¦š¸ÉÁ®¤µ³­¤˜ n°„µ¦´„œÎµ„µ¦Á„—¥°—šª· ¸‡¼–‹µ„˜oœ „¨oª¥Å¤o Paphiopedilum ‘Delrosi’ š¸É˜´—­nªœ¥°—°°„...... 33 «¹„¬µ­¼˜¦°µ®µ¦š¸ÉÁ®¤µ³­¤˜n°„µ¦´„œÎµ„µ¦Á„—¥°—‹µ„­· nªœ o° °Š˜oœ „¨oª¥Å¤o Paphiopedilum ‘Delrosi’ ...... 36 «¹„¬µ­¼˜¦°µ®µ¦š¸ÉÁ®¤µ³­¤˜n°„µ¦´„œÎµ„µ¦Á„—‡· ¨¨´­ ®¦º° somatic embryos ‹µ„œÁœ·Ê ºÊ°Á¥ºÉ°¦³®ªnµŠ o° °Š„¨oª¥Å¤ o Paphiopedilum ‘Delrosi’ ...... 39 «¹„¬µ­¼˜¦°µ®µ¦š¸ÉÁ®¤µ³­¤˜n°„µ¦´„œÎµ„µ¦Á„—‡¨¨· ´­ ®¦º° somatic embryos ‹µ„œ­·Ê nªœÄ °Š„¨oª¥Å¤o Paphiopedilum ‘Delrosi’...... 42 4 Ÿ¨„µ¦š—¨°Š...... 45 «¹„¬µ­¼˜¦°µ®µ¦š¸ÉÁ®¤µ³­¤˜°„µ¦n ´„œÎµ„µ¦Á„·—¥°—šª¸‡¼–‹µ„­ªœn ¥°— °Š„¨oª¥Å¤ o Paphiopedilum ‘Delrosi’...... 45

Ž šš¸É ®œoµ «¹„¬µŸ¨ °Š­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ TDZ BA Kn 2,4-D ¨³ Ads ˜n°„µ¦´„ œÎµ„µ¦Á„·—¥°—šª¸‡¼–‹µ„­ªœ¥°— °Š„¨n oª¥Å¤ o Paphiopedilum ‘Delrosi’ ...... 45 «¹„¬µŸ¨ °Š­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ TDZ BA Kn ¨³ 2,4-D ˜n°„µ¦´„ œÎµ„µ¦Á„·—¥°—šª¸‡¼–‹µ„­ªœ¥°— °Š„¨n oª¥Å¤ o Paphiopedilum ‘Delrosi’ ...... 51 «¹„¬µœ— °Šœ· Ê嘵¨š¸ÉÁ®¤µ³­¤˜n°„µ¦´„œÎµ„µ¦Á„—¥°—šª· ¸‡¼–‹µ„­ªœn ¥°— °Š„¨oª¥Å¤ o Paphiopedilum ‘Delrosi’...... 55 «¹„¬µ­¼˜¦°µ®µ¦š¸ÉÁ®¤µ³­¤˜°„µ¦n ´„œÎµ„µ¦Á„·—¥°—šª¸‡¼–‹µ„˜œ„¨o oª¥Å¤ o Paphiopedilum ‘Delrosi’ š¸É˜´—­nªœ¥°—°°„...... 60 «¹„¬µ­¼˜¦°µ®µ¦š¸ÉÁ®¤µ³­¤˜°„µ¦n ´„œÎµ„µ¦Á„·—¥°—‹µ„­nªœ o° °Š„¨oª¥Å¤ o

Paphiopedilum ‘Delrosi’...... 64 «¹„¬µ­¼˜¦°µ®µ¦š¸ÉÁ®¤µ³­¤˜°„µ¦n ´„œÎµ„µ¦Á„·—‡¨¨´­ ®¦º° somatic embryos ‹µ„·ÊœÁœºÊ°Á¥ºÉ°¦³®ªnµŠ o° °Š„¨oª¥Å¤ o Paphiopedilum ‘Delrosi’. 70 «¹„¬µ­¼˜¦°µ®µ¦š¸ÉÁ®¤µ³­¤˜°„µ¦n ´„œÎµ„µ¦Á„·—‡¨¨´­ ®¦º° somatic

embryos ‹µ„·Êœ­nªœÄ °Š„¨oª¥Å¤ o Paphiopedilum ‘Delrosi’...... 72

5 °£·ž¦µ¥Ÿ¨„µ¦š—¨°Š...... 74 «¹„¬µ­¼˜¦°µ®µ¦š¸ÉÁ®¤µ³­¤˜°„µ¦n ´„œÎµ„µ¦Á„·—¥°—šª¸‡¼–‹µ„­ªœ¥°— °Šn „¨oª¥Å¤ o Paphiopedilum ‘Delrosi’ ...... 74 «¹„¬µŸ¨ °Š­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ TDZ, BA, Kn, 2,4-D ¨³ Ads ˜n° „µ¦´„œÎµ„µ¦Á„·—¥°—šª¸‡¼–‹µ„­nªœ¥°— °Š„¨oª¥Å¤ o Paphiopedilum ‘Delrosi’ ...... 74 «¹„¬µŸ¨ °Š­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ TDZ BA Kn ¨³ 2,4-D ˜n°„µ¦´„ œÎµ„µ¦Á„·—¥°—šª¸‡¼–‹µ„­ªœ¥°— °Š„¨n oª¥Å¤ o Paphiopedilum ‘Delrosi’ ...... 75 «¹„¬µœ— °Šœ· Ê嘵¨š¸ÉÁ®¤µ³­¤˜n°„µ¦´„œÎµ„µ¦Á„—¥°—šª· ¸‡¼–‹µ„­ªœn ¥°— °Š„¨oª¥Å¤ o Paphiopedilum ‘Delrosi’...... 76 «¹„¬µ­˜¦°µ®µ¦š¼ ¸ÉÁ®¤µ³­¤˜°„µ¦n ´„œÎµ„µ¦Á„·—¥°—šª¸‡¼–‹µ„˜œ„¨o oª¥Å¤ o Paphiopedilum ‘Delrosi’ š¸É˜´—­nªœ¥°—°°„...... 77

 šš¸É ®œoµ «¹„¬µ­¼˜¦°µ®µ¦š¸ÉÁ®¤µ³­¤˜°„µ¦n ´„œÎµ„µ¦Á„·—¥°—‹µ„­nªœ o° °Š„¨oª¥Å¤o Paphiopedilum ‘Delrosi’...... 78 «¹„¬µ­¼˜¦°µ®µ¦š¸ÉÁ®¤µ³­¤˜°„µ¦n ´„œÎµ„µ¦Á„·—‡¨¨´­ ®¦º° somatic embryos ‹µ„·ÊœÁœºÊ°Á¥ºÉ°¦³®ªnµŠ o° °Š„¨oª¥Å¤ o Paphiopedilum ‘Delrosi’. 78 «¹„¬µ­¼˜¦°µ®µ¦š¸ÉÁ®¤µ³­¤˜°„µ¦n ´„œÎµ„µ¦Á„·—‡¨¨´­ ®¦º° somatic embryos ‹µ„·Êœ­nªœÄ °Š„¨oª¥Å¤ o Paphiopedilum ‘Delrosi’...... 79 6 ­¦»žŸ¨„µ¦š—¨°Š...... 81 «¹„¬µ­¼˜¦°µ®µ¦š¸ÉÁ®¤µ³­¤˜°„µ¦n ´„œÎµ„µ¦Á„·—¥°—šª¸‡¼–‹µ„­ªœ¥°— °Šn „¨oª¥Å¤ o Paphiopedilum ‘Delrosi’ ...... 81 «¹„¬µŸ¨ °Š­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ TDZ, BA, Kn, 2,4-D ¨³ Ads ˜n°„µ¦´„œÎµ„µ¦Á„—¥°—š· ª¸‡¼–‹µ„­nªœ¥°— °Š„¨oª¥Å¤ o

Paphiopedilum ‘Delrosi’...... 81 «¹„¬µŸ¨ °Š­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ TDZ BA Kn ¨³ 2,4-D ˜n°„µ¦ ´„œÎµ„µ¦Á„·—¥°—šª¸‡¼–‹µ„­nªœ¥°— °Š„¨ª¥Å¤o o Paphiopedilum ‘Delrosi’ ...... 81

«¹„¬µœ— °Šœ· Ê嘵¨š¸ÉÁ®¤µ³­¤˜n°„µ¦´„œÎµ„µ¦Á„—¥°—šª· ¸‡¼–‹µ„­ªœn

¥°— °Š„¨oª¥Å¤ o Paphiopedilum ‘Delrosi’...... 82 «¹„¬µ­¼˜¦°µ®µ¦š¸ÉÁ®¤µ³­¤˜°„µ¦n ´„œÎµ„µ¦Á„·—¥°—šª¸‡¼–‹µ„˜œ„¨o oª¥Å¤ o Paphiopedilum ‘Delrosi’ š¸É˜´—­nªœ¥°—°°„...... 82 «¹„¬µ­¼˜¦°µ®µ¦š¸ÉÁ®¤µ³­¤˜°„µ¦n ´„œÎµ„µ¦Á„·—¥°—‹µ„­nªœ o° °Š„¨oª¥Å¤ o Paphiopedilum ‘Delrosi’...... 83 «¹„¬µ­¼˜¦°µ®µ¦š¸ÉÁ®¤µ³­¤˜°„µ¦n ´„œÎµ„µ¦Á„·—‡¨¨´­ ®¦º° somatic embryos ‹µ„·ÊœÁœºÊ°Á¥ºÉ°¦³®ªnµŠ o° °Š„¨oª¥Å¤ o Paphiopedilum ‘Delrosi’ 83 «¹„¬µ­¼˜¦°µ®µ¦š¸ÉÁ®¤µ³­¤˜°„µ¦n ´„œÎµ„µ¦Á„·—‡¨¨´­ ®¦º° somatic embryos ‹µ„·Êœ­nªœÄ °Š„¨oª¥Å¤ o Paphiopedilum ‘Delrosi’...... 83 o°Á­œ°Âœ³...... 85 ¦¦–µœ»„¦¤...... 86 £µ‡Ÿœª„...... 91 ‡Îµ°›·µ¥‡Îµ¥n°...... 92

 šš¸É ®œoµ £µ‡Ÿœª„°Š‡rž¦³„° °Š­¼˜¦°µ®µ¦ Murashige and Skoog...... 93 ž¦³ª´˜·Ÿ¼oª·‹´¥...... 94

‘ ­µ¦´˜µ¦µŠ ˜µ¦µŠš¸É ®œoµ 1 ­µ¦‡ª‡»¤„µ¦Á˜Ã˜· TDZ, BA, Kn, 2,4-D ¨³ Ads ˜n°„µ¦´„œÎµ„µ¦Á„·— ¥°—šª¸‡¼–‹µ„­ªœ¥°— °Š„¨n oª¥Å¤ o Paphiopedilum ‘Delrosi’ ...... 27 2 ­µ¦‡ª‡»¤„µ¦Á˜Ã˜· TDZ, BA, Kn ¨³ 2,4-D ˜n°„µ¦´„œÎµ„µ¦Á„—¥°—· šª¸‡¼–‹µ„­nªœ¥°— °Š„¨oª¥Å¤ o Paphiopedilum ‘Delrosi’...... 30 3 œ·— °ŠœÊ嘵¨Â¨³­µ¦‡ª‡»¤„µ¦Á˜·Ã˜š¸ÉčoĜ„µ¦´„œÎµ„µ¦Á„·—¥°— šª¸‡¼–‹µ„­nªœ¥°— °Š„¨oª¥Å¤ o Paphiopedilum ‘Delrosi’...... 32 4 œ·— °ŠœÊ嘵¨Â¨³­µ¦‡ª‡»¤„µ¦Á˜·Ã˜š¸ÉčoĜ„µ¦´„œÎµ„µ¦Á„·—¥°— šª¸‡¼–‹µ„˜oœ„¨ª¥Å¤o o Paphiopedilum ‘Delrosi’ š¸É˜´—¥°—°°„ ...... 34 5 œ·— °ŠœÊ嘵¨Â¨³­µ¦‡ª‡»¤„µ¦Á˜·Ã˜š¸ÉčoĜ„µ¦´„œÎµ„µ¦Á„·—¥°—‹µ„ ­nªœ o° °Š˜oœ„¨oª¥Å¤ o Paphiopedilum ‘Delrosi’...... 37

6 œ·— °ŠœÊ嘵¨Â¨³­µ¦‡ª‡»¤„µ¦Á˜·Ã˜š¸ÉčoĜ„µ¦´„œÎµ„µ¦Á„·—‡¨¨´­ ®¦º° somatic embryos ‹µ„·ÊœÁœºÊ°Á¥ºÉ°¦³®ªnµŠ o° °Š„¨oª¥Å¤ o Paphiopedilum‘Delrosi’...... 40 7 œ·— °ŠœÊ嘵¨Â¨³­µ¦‡ª‡»¤„µ¦Á˜·Ã˜š¸ÉčoĜ„µ¦´„œÎµ„µ¦Á„·—‡¨¨´­

®¦º° somatic embryos ‹µ„·Êœ­nªœÄ °Š„¨oª¥Å¤ o Paphiopedilum

‘Delrosi’...... 43 8 Ÿ¨ °Š­¼˜¦°µ®µ¦—´—ž¨Š Hyponex ¦nª¤„´­µ¦‡ª‡»¤„µ¦Á˜·Ã˜TDZ, BA, Kn, 2,4-D ¨³ Ads ˜n°„µ¦´„œÎµ„µ¦Á„·—¥°—šª¸‡¼–‹µ„­ªœ¥°— °Šn „¨oª¥Å¤ o Paphiopedilum ‘Delrosi’ ®¨´Š‹µ„„µ¦Á¡µ³Á¨¸Ê¥ŠÁžœ¦³¥³Áª¨µ} 16 ­´ž—µ®r ...... 47 9 Ÿ¨ °Š­¼˜¦°µ®µ¦—´—ž¨Š Hyponex ¦nª¤„´­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ TDZ, BA, Kn, 2,4-D ¨³ Ads ˜n°„µ¦Á‹¦·Á˜·Ã˜ °Š­nªœ¥°—Á¦·É¤˜œ °Šo „¨oª¥Å¤ o Paphiopedilum ‘Delrosi’ ®¨´Š‹µ„„µ¦Á¡µ³Á¨¸Ê¥ŠÁžœ¦³¥³Áª¨µ} 16 ­´ž—µ®r ...... 48 10 Ÿ¨ °Š­¼˜¦°µ®µ¦—´—ž¨Š Hyponex ¦nª¤„´­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ TDZ, BA, Kn ¨³ 2,4-D ˜n°„µ¦´„œÎµ„µ¦Á„·—¥°—šª¸‡¼–‹µ„­nªœ¥°— °Š „¨oª¥Å¤ o Paphiopedilum ‘Delrosi’ ®¨´Š‹µ„„µ¦Á¡µ³Á¨¸Ê¥ŠÁžœ¦³¥³Áª¨µ} 16 ­´ž—µ®r ...... 52

’ ˜µ¦µŠš¸É ®œoµ 11 Ÿ¨ °Š­¼˜¦°µ®µ¦—´—ž¨Š Hyponex ¦nª¤„´­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ TDZ, BA, Kn ¨³ 2,4-D ˜n°„µ¦Á‹¦·Á˜·Ã˜ °Š¥°—Á¦·É¤˜oœ °Š„¨oª¥Å¤o Paphiopedilum ‘Delrosi’ ®¨´Š‹µ„„µ¦Á¡µ³Á¨¸Ê¥ŠÁžœ¦³¥³Áª¨µ} 16 ­´ž—µ®r ...... 53 12 Ÿ¨ °Š­¼˜¦°µ®µ¦—´—ž¨Š Hyponex ¦nª¤„´œÊ嘵¨ 3 œ·— ¨³­µ¦‡ª‡»¤ „µ¦Á˜·Ã˜ TDZ, Kn ¨³ 2,4-D ˜n°„µ¦´„œÎµ„µ¦Á„·—¥°—šª¸‡¼–‹µ„ ­nªœ¥°— °Š„¨oª¥Å¤ o Paphiopedilum ‘Delrosi’ ®¨´Š‹µ„„µ¦Á¡µ³Á¨¸Ê¥Š Áž}œ¦³¥³Áª¨µ 16 ­´ž—µ®r ...... 57 13 Ÿ¨ °Š­¼˜¦°µ®µ¦—´—ž¨Š Hyponex ¦nª¤„´œÊ嘵¨ 3 œ·— ¨³­µ¦‡ª‡»¤ „µ¦Á˜·Ã˜ TDZ, Kn ¨³ 2,4-D ˜n°„µ¦Á‹¦·Á˜·Ã˜ °Š¥°—Á¦·É¤˜oœ °Š „¨oª¥Å¤ o Paphiopedilum ‘Delrosi’ ®¨´Š‹µ„„µ¦Á¡µ³Á¨¸Ê¥ŠÁžœ¦³¥³Áª¨µ}

16 ­´ž—µ®r ...... 58 14 Ÿ¨ °Š­¼˜¦°µ®µ¦—´—ž¨Š Hyponex ¦nª¤„´œÊ嘵¨ 3 œ·— ¨³­µ¦‡ª‡»¤ „µ¦Á˜·Ã˜ TDZ, Kn ¨³ 2,4-D ˜n°„µ¦´„œÎµ„µ¦Á„·—¥°—šª¸‡¼–‹µ„˜oœ „¨oª¥Å¤ o Paphiopedilum ‘Delrosi’ š¸É˜´—­nªœ¥°—°°„ ®¨´Š‹µ„„µ¦Á¡µ³

Á¨¸Ê¥ŠÁž}œ¦³¥³Áª¨µ 16 ­´ž—µ®r ...... 61

15 Ÿ¨ °Š­¼˜¦°µ®µ¦—´—ž¨Š Hyponex ¦nª¤„´œÊ嘵¨ 3 œ·— ¨³­µ¦‡ª‡»¤ „µ¦Á˜·Ã˜ TDZ, Kn ¨³ 2,4-D ˜n°„µ¦´„œÎµ„µ¦Á„·—¥°—‹µ„­ªœ n o° °Š„¨oª¥Å¤ o Paphiopedilum ‘Delrosi’ ®¨´Š‹µ„„µ¦Á¡µ³Á¨¸Ê¥ŠÁžœ¦³¥³} Áª¨µ 16 ­´ž—µ®r...... 66

“ ­µ¦´£µ¡ £µ¡š ¸É ®œoµ 1 ¨´„¬–³šµŠ¡§„¬«µ­˜¦r °Š„¨oª¥Å¤o¦°ŠÁšoµœµ¦¸ (Paphiopedilum) ...... 7 2 ˜oœ„¨oª¥Å¤o Paphiopedilum ‘Delrosi’ Ĝ ª—ž¨°—ÁºÊ° °µ¥ » 14 Á—º°œ®¨´Š‹µ„ Á¡µ³Á¤¨È—...... 25 3 ¨´„¬–³ °Š„¨oª¥Å¤o Paphiopedilum ‘Delrosi’ ...... 28 4 ¨´„¬–³˜oœ„¨ª¥Å¤o o Paphiopedilum ‘Delrosi’ ®¨´Š‹µ„Á¡µ³Á¨¸Ê¥ŠÄœš¸É¤º—..... 35 5 ¨´„¬–³„µ¦Á¡µ³Á¨¸Ê¥Š˜oœ„¨oª¥Å¤o Paphiopedilum ‘Delrosi’ š¸É˜´—­nªœ¥°— °°„...... 35 6 ¨´„¬–³„µ¦Á¡µ³Á¨¸Ê¥Š­nªœ o° °Š˜oœ„¨oª¥Å¤ o Paphiopedilum ‘Delrosi’...... 38 7 ¨´„¬–³„µ¦Á¡µ³Á¨¸Ê¥ŠÁœºÊ°Á¥ºÉ°¦³®ªnµŠ o° °Š„¨oª¥Å¤o Paphiopedilum ‘Delrosi’...... 41

8 ¨´„¬–³„µ¦Á¡µ³Á¨¸Ê¥Š·Êœ­nªœÄ °Š˜oœ„¨oª¥Å¤o Paphiopedilum ‘Delrosi’ .. 44 9 Ÿ¨ °Š­¼˜¦°µ®µ¦—´—ž¨Š Hyponex ¦nª¤„´­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ TDZ, BA, Kn, 2,4-D ¨³ Ads ˜n°„µ¦Á‹¦·Á˜·Ã˜Â¨³„µ¦´„œÎµ„µ¦Á„·—¥°— šª¸‡¼–‹µ„­nªœ¥°— °Š„¨oª¥Å¤o Paphiopedilum ‘Delrosi’ ®¨´Š‹µ„„µ¦

Á¡µ³Á¨¸Ê¥ŠÁž}œ¦³¥³Áª¨µ 16 ­´ž—µ®r...... 49

10 Ÿ¨ °Š­¼˜¦°µ®µ¦—´—ž¨Š Hyponex ¦nª¤„´­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ TDZ, BA, Kn ¨³ 2,4-D ˜n°„µ¦Á‹¦·Á˜·Ã˜Â¨³„µ¦´„œµ„µ¦Á„Î ·—¥°—šª¸‡¼– ‹µ„­nªœ¥°— °Š„¨oª¥Å¤o Paphiopedilum ‘Delrosi’ ®¨´Š‹µ„„µ¦Á¡µ³ Á¨¸Ê¥ŠÁž}œ¦³¥³Áª¨µ 16 ­´ž—µ®r ...... 54 11 Ÿ¨ °Š­¼˜¦°µ®µ¦—´—ž¨Š Hyponex ¦nª¤„´œÊ嘵¨ 3 œ·— ¨³­µ¦‡ª‡»¤ „µ¦Á˜·Ã˜ TDZ, Kn ¨³ 2,4-D ˜n°„µ¦Á‹¦·Á˜·Ã˜Â¨³„µ¦´„œµ„µ¦Î Á„·—¥°—šª¸‡¼–‹µ„­nªœ¥°— °Š„¨ª¥Å¤o o Paphiopedilum ‘Delrosi’ ®¨´Š ‹µ„„µ¦Á¡µ³Á¨¸Ê¥ŠÁžœ¦³¥³Áª¨µ} 16 ­´ž—µ®r...... 59 12 Ÿ¨ °Š­¼˜¦°µ®µ¦—´—ž¨Š Hyponex ¦nª¤„´œÊ嘵¨ 3 œ·— ¨³­µ¦‡ª‡»¤ „µ¦Á˜·Ã˜ TDZ, Kn ¨³ 2,4-D ˜n°„µ¦´„œÎµ„µ¦Á„·—¥°—šª¸‡¼–‹µ„˜oœ „¨oª¥Å¤ o Paphiopedilum ‘Delrosi’ š¸É˜´—­nªœ¥°—°°„ ®¨´Š‹µ„„µ¦Á¡µ³ Á¨¸Ê¥ŠÁž}œ¦³¥³Áª¨µ 16 ­´ž—µ®r ...... 62

” £µ¡š ¸É ®œoµ 13 Ÿ¨ °Š­¼˜¦°µ®µ¦—´—ž¨Š Hyponex ¦nª¤„´œÊ嘵¨ 3 œ·— ¨³­µ¦‡ª‡»¤ „µ¦Á˜·Ã˜ TDZ, Kn ¨³ 2,4-D ˜n°„µ¦´„œÎµ„µ¦Á„·—¥°—‹µ„­ªœ n o° °Š„¨oª¥Å¤o Paphiopedilum ‘Delrosi’ ®¨´Š‹µ„„µ¦Á¡µ³Á¨¸Ê¥ŠÁžœ¦³¥³} Áª¨µ 16 ­´ž—µ®r...... 67 14 Ÿ¨ °Š­¼˜¦°µ®µ¦—´—ž¨Š Hyponex ¦nª¤„´œÊ嘵¨ 3 œ·— ¨³­µ¦‡ª‡»¤ „µ¦Á˜·Ã˜ TDZ ¨³ 2,4-D ˜n°„µ¦´„œÎµ„µ¦Á„·—‡¨¨´­ ®¦º° somatic embryos‹µ„·ÊœÁœºÊ°Á¥ºÉ°¦³®ªnµŠ o° °Š„¨oª¥Å¤ o Paphiopedilum ‘Delrosi’ ...... 71 15 Ÿ¨ °Š­¼˜¦°µ®µ¦—´—ž¨Š Hyponex ¦nª¤„´œÊ嘵¨ 3 œ·— ¨³­µ¦‡ª‡»¤ „µ¦Á˜·Ã˜ TDZ ¨³ 2,4-D ˜n°„µ¦´„œÎµ„µ¦Á„·—‡¨¨´­ ®¦º° somatic embryos‹µ„·Êœ­nªœÄ °Š„¨oª¥Å¤ o Paphiopedilum ‘Delrosi’...... 73

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3. Á¡ºÉ°«¹„¬µ­˜¦°µ®µ¦š¼ ¸ÉÁ®¤µ³­¤˜n°„µ¦´„œÎµ„µ¦Á„·—‡¨¨´­ ®¦º° somatic embryos

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3. °Á ˜„µ¦«¹„¬µ 1. «¹„¬µ­¼˜¦°µ®µ¦š¸ÉÁ®¤µ³­¤˜n°„µ¦´„œÎµ„µ¦Á„—¥°—šª· ¸‡¼–‹µ„­ªœ¥°—n ˜oœš¸É˜´— ­nªœ¥°—°°„ ¨³ o° °Š„¨oª¥Å¤ o Paphiopedilum ‘Delrosi’ ‹µ„­¼˜¦°µ®µ¦—´—ž¨Š Hyponex š¸É ž¦³„°—ª¥­µ¦‡ª‡o »¤„µ¦Á˜·Ã˜ TDZ, BA, Kn, 2,4-D ¨³ Ads Áž}œ¦³¥³Áª¨µ 16 ­´ž—µ® r 2. «¹„¬µœ·— °ŠœÊ嘵¨š¸ÉÁ®¤µ³­¤˜n°„µ¦´„œÎµ„µ¦Á„—¥°—š· ª¸‡¼–‹µ„­nªœ¥°— ˜oœš¸É ˜´—­nªœ¥°—°°„ ¨³ o° °Š„¨oª¥Å¤ o Paphiopedilum ‘Delrosi’ ‹µ„­¼˜¦°µ®µ¦—´—ž¨Š Hyponex š¸Éž¦³„°—ª¥œo Ê嘵¨Ž¼Ã‡¦­ „¨¼Ã‡­ ¨³¤°¨Ã˜­ ‡ªµ¤Á o¤ oœ 20 „¦´¤˜n°¨·˜¦ Áž}œ¦³¥³Áª¨µ 16 ­´ž—µ® r 3

3. «¹„¬µœ·— °ŠœÊ嘵¨Â¨³­¼˜¦°µ®µ¦š¸ÉÁ®¤µ³­¤˜n°„µ¦´„œÎµ„µ¦Á„—‡¨¨· ´­ ®¦º° somatic embryos ‹µ„ÁœºÊ°Á¥ºÉ°¦³®ªnµŠ o° ¨³·Êœ­nªœÄ„¨oª¥Å¤ o Paphiopedilum ‘Delrosi’ ‹µ„ ­¼˜¦°µ®µ¦ 1/2 MS (1/2 strength macro-and full-strength micro elements) š¸Éž¦³„°—oª¥œÊ嘵¨ Ž¼Ã‡¦­ „¨¼Ã‡­ ¨³¤°¨Ã˜­ ‡ªµ¤Á o¤ oœ 20 „¦´¤˜n°¨·˜¦ ¦nª¤„´­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ TDZ, Kn ¨³ 2,4-D Áž}œ¦³¥³Áª¨µ 16 ­´ž—µ® r

4. ž¦³Ã¥œrš¸É‡µ—ªnµ‹³Å—o¦´ 1. š¦µ™¹Š­¼˜¦°µ®µ¦ ¨³œ·— °ŠœÊ嘵¨š¸ÉÁ®¤µ³­¤˜°„µ¦n ´„œÎµ„µ¦Á„·—¥°— šª¸‡¼–‹µ„­nªœ¥°— ˜oœš¸É˜´—­nªœ¥°—°°„ ¨³ o° °Š„¨oª¥Å¤ o Paphiopedilum ‘Delrosi’ 2. š¦µ™¹Š­¼˜¦°µ®µ¦ ¨³œ·— °ŠœÊ嘵¨š¸ÉÁ®¤µ³­¤˜n°„µ¦´„œÎµ„µ¦Á„·—‡¨¨´­ ®¦º° somatic embryos ‹µ„·ÊœÁœºÊ°Á¥ºÉ°¦³®ªnµŠ o° ¨³·Êœ­nªœÄ„¨oª¥Å¤ o Paphiopedilum ‘Delrosi’

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„¨oª¥Å¤o (orchid) Áž}œÅ¤o—°„Äœ„¨»n¤¡ºÄÁ¨¸Ê¥ŠÁ—¥ª¸É (monocotyledon) ‹´—°¥¼nĜªŠ«r „¨oª¥Å¤o®¦º°ªŠ«r°°¦r‡·Á—Ž·°¸ (Family Orchidaceae) Áž}œ¡ºªŠ«Ä®r nš¸É­»—Äœ¦¦—µ¡¦¦–¡ºš´Éª è„ ž{‹‹»´œ¤¸°¥¼nž¦³¤µ– 796 ­„»¨ (Genus) ¨³¤µ„„ªnµ 17,500 œ·— (Species) ¨³œ´Ážœ¡} º ªŠ«rÄ®nš¸É­»—Äœž¦³Áš«Åš¥ ×¥¤¸„µ¦­Îµ¦ª‹¡Â¨oªšŠ­´Ê ·Êœ 168 ­„»¨ ¨³¤µ„„ªnµ 1,170 œ·— ®¦º°Áž}œ®œ¹ÉŠÄœ­·®oµ °Š„¨oª¥Å¤oš¸É¡šªÃ¨„´É œ°„‹µ„œ¸Ê „¨oª¥Å¤Åš¥¥o ´Š¤¸‡ªµ¤®¨µ„®¨µ¥š´ÊŠ ™·Éœ“µœš¸É°¥Â¨³¡¼n ´œ›»„¦¦¤Å¤nœo°¥Åž„ªnµž¦³Áš«Á ˜¦°œ°o ºÉœÇ (¦³¡ ¸ 2516; ­¨·¨ 2549)

1. ž¦³ª´˜·Â¨³ª·ª´•œµ„µ¦ °Š„¨oª¥Å¤oĜž¦³Áš«Åš¥ ˜nÁ—·¤Äœž¦³Áš«Åš¥œœ¥´Ê ´ŠÅ¤n¤¸Ÿ¼oėĮo‡ªµ¤­Îµ‡´„´„¨oª¥Å¤o¤µ„œ´„ ‹œ¤µ™¹Š˜oœ ­¤´¥¦´˜œÃ„­·œš¦r‹¹Š¤¸Ÿ¼o¦·Á¦·É¤ž¨¼„Á¨¸Ê¥Š„¨ª¥Å¤o o ¹Êœ ×¥œµ¥Á±œ¦¸É °µ¨µµ­Á˜°¦r (˜oœ˜¦³„¼¨ “Á«ª˜

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2. „¨oª¥Å¤o¦°ŠÁšoµœµ¦ ¸ „¨oª¥Å¤o¦°ŠÁšoµœµ¦¸ ®¦º°š¸É£µ¬µ°´Š„§¬Á¦¸¥„ªnµ Lady’s Slipper œ´Êœ ¤¸™·Éœ„εÁœ·—š´ÊŠÄœ Á ˜¦o°œÂ¨³®œµª °ŠÃ¨„ Ášnµš¸É¡Â¨oªšªÃ¨„¤´É ¸ 4 ­„»¨ 125 œ·— ‡º° ­„»¨ Cypripedium ¤¸ 35 4 5

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3. „¨oª¥Å¤o¦°ŠÁšoµœµ¦¸­„»¨ Paphiopedilum 3.1 „µ¦„¦³‹µ¥¡´œ› »r „¨oª¥Å¤o¦°ŠÁšoµœµ¦¸­„»¨ Paphiopedilum ‹´—Ážœ¡} ´œ›»rŤoš¸É¤¸™·Éœ„εÁœ—ÄœÁ ˜¦· o°œ „¦³‹µ¥¡´œ›»r°¥¼n˜µ¤›¦¦¤µ˜·œ´‹µ„ÂœªÁšº°„Á µ®·¤µ¨¥¨Š¤µ­´ ¼n˜°œ¨nµŠ ×¥ÁŒ¡µ³Â™Á°Á¥˜³¸ ª´œ°°„ÁŒ¥ŠÄ˜¸ o (¦ª¤š´ÊŠÅš¥) °·œÁ—¸¥ °·œÃ—œ¸ÁŽ¸¥ £µ‡˜³ªœ°°„ÁŒ´ ¸¥ŠÄ˜o °Š‹¸œ œ·ª„·œ¸ ¢d¨·žždœ­r ¨³®¤¼nÁ„µ³ÃŽÃ¨¤°œ ˜n‡œÄœ¡œšºÊ ¸ÉŤnœ·¥¤œÎµ¤µž¨„Á¨¼ ¸Ê¥Š„´œ ž¨n°¥Ä®o°¥¼n˜µ¤›¦¦¤µ˜· ¨³ µŠœµ˜·¥´Š¤¸‡ªµ¤ÁºÉ°ªnµºÉ° °Š„¨oª¥Å¤o¦°ŠÁšoµœµ¦¸Áž}œºÉ°š¸ÉŤnÁžœ¤Š‡¨} (¦³¡¸ 2535; °»Å¦ 2549) 3.2 „¨oª¥Å¤o¦°ŠÁšoµœµ¦¸Äœž¦³Áš«Åš¥

„¨oª¥Å¤o¦°ŠÁšoµœµ¦¸š¸É¡ªnµ¤¸™·Éœ„εÁœ·—°¥¼nĜž¦³Áš«Åš¥Áž}œ„¨oª¥Å¤¦°ŠÁšo oµœµ¦¸ ­„»¨ Paphiopedilum ž{‹‹»´œš¸É‡oœ¡Â¨ª¤o ¸š´ÊŠ®¤— 17 œ·— ‡º° (°»Å¦ 2549; ›µ¦š·¡¥r 2010) 1. ¦°ŠÁšoµœµ¦¸‡µŠ„‡°Â—Š (Paph. appletonianum var. wolterianum) 2. ¦°ŠÁšoµœµ¦¸¤nªŠ­Š ¨µ ®¦º°¦°ŠÁšoµœµ¦¸‡µŠ„£µ‡Ä˜o (Paph. barbatum)

3. ¦°ŠÁšoµœµ¦¸ µ®°¥ (Paph. bellatulum)

4. ¦°ŠÁšoµœµ¦¸‡µŠ„ ®¦º°¦°ŠÁš oµœµ¦¸Åš¥Â¨œ—r (Paph. callosum) 5. ¦°ŠÁšoµœµ¦¸—°¥˜»Š (Paph. charlesworthii) 6. ¦°ŠÁšoµœµ¦¸Á®¨º°Šž¦µ‹¸œ ®¦º°¦°ŠÁšoµœµ¦¸Á®¨º°Š„µ‹œr ®¦º°¦°ŠÁšoµœµ¦¸Á®¨º°Š °»—¦ (Paph. concolor) 7. ¦°ŠÁšoµœµ¦¸Á®¨º°Š„¦³¸É (Paph. exul) 8. ¦°ŠÁšoµœµ¦¸ µª»¤¡¦ (Paph. godefroyae) 9. ¦°ŠÁšoµœµ¦¸Á®¨º°Š˜¦´Š ®¦º°¦°ŠÁšoµœµ¦¸Á®¨º°Š¡´ŠŠµ (Paph. godefroyae var. leucochilum) 10. ¦°ŠÁšoµœµ¦¸Á®¨º°ŠÁ¨¥ (Paph. hirsutissimum var. esquirolei) 11. ¦°ŠÁšoµœµ¦¸°·œŽ·„Áœn (Paph. insigne) 12. ¦°ŠÁšoµœµ¦¸ µª­˜¼¨ (Paph. niveum) 13. ¦°ŠÁšoµœµ¦¸Á¤º°Š„µ‹œr ®¦º°¦°ŠÁšoµœµ¦¸Á¸¥Š—µª (Paph. parishii) 14. ¦°ŠÁšoµœµ¦¸že„¤¨Šž° ®¦º°¦°ŠÁšoµœµ¦¸­» ³„¼¨ (Paph. sukhakulii) 6

15. ¦°ŠÁšoµœµ¦¸°·œšœœšr (Paph.villosum) 16. ¦°ŠÁšoµœµ¦¸n°Š°nµŠš°Š (Paph. x Ang Thong) 17. ¦°ŠÁšoµœµ¦¸Á„µ³oµŠ (Paph. x Siamensis)

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7.1.1. ­µ¦°œ·œš¦¸¥r (inorganic component) ­¼˜¦°µ®µ¦­Îµ®¦´Á¡µ³Á¨¥ŠÁœ¸Ê ºÊ°Á¥ºÉ°¡ºš»„ ­¼˜¦˜o°Š¤¸Á„¨º°°œ·œš¦¸¥rÁžœ°Š‡} rž¦³„°¡ºÊœ“µœ Á¡ºÉ°ÄoĜ„µ¦Á‹¦Á˜· ·Ã˜ÁnœÁ—¸¥ª„´¡ºš¸Éž¨¼„ Ĝ›¦¦¤µ˜· ץnŠÁž}œ 2 „¨»n¤ ‡º° 7.1.1.1 ›µ˜»°µ®µ¦š¸É¡º˜o°Š„µ¦Äœž¦·¤µ–¤µ„ (macroelements) ŗo„ n C H O N P K S Ca ¨³ Mg Ž¹ÉŠ¤´„¦ª¤Åž™¹Š chlorine ¨³ sodium —oª¥ ­¼˜¦°µ®µ¦¡ºÊœ“µœ (basal medium) š´ÉªÅž‹³¤¸„µ¦Á˜·¤ macroelements ¨ŠÅžÄœž¦·¤µ–š¸É¤µ„„ªnµ 30 ppm (mg/l) ×¥›µ˜»°µ®µ¦˜nµŠÇ Á®¨nµœ¸ÊÁž}œ­·ÉŠ‹ÎµÁž}œ˜n°„µ¦Á‹¦·Á˜·Ã˜ °ŠÁŽ¨¨r¨³Áœ°Á¥ºÊ ºÉ°¡º 7.1.1.2 ›µ˜»°µ®µ¦š¸É¡º˜o°Š„µ¦Äœž¦·¤µ–œo°¥ (microelements) ŗ„o n Fe Cl Mn Cu Zn B ¨³ Mo ×¥›µ˜»°µ®µ¦Á®¨nµœ¸Ê‹³šÎµ®œoµš¸ÉÁž}œ co-factor ¨³ inducer Ĝ„¦³ªœ„µ¦ ­´ŠÁ‡¦µ³®r °ŠÁ°œÅŽ¤r ¨³Äœ°µ®µ¦­˜¦¡¼ ºÊœ“µœšªÅž‹³¤´É „µ¦Á˜¸ ·¤ microelements ¨ŠÅžÄœ ž¦·¤µ–š¸Éœo°¥„ªnµ 30 ppm (mg/l) 7.1.2. ­µ¦°·œš¦¸¥r (organic component) ‡º° ­µ¦š¸É¤¸°Š‡rž¦³„° °Š‡µ¦r°œ (C) űצ

Á‹œ (H) ¨³°°„Ž·Á‹œ (O) ŗo„ n 7.1.2.1 ‡µ¦r°œ (carbon) ×¥ž„˜Â¨· oªÁŽ¨¨r¡º­¸Á ¸¥ªÁ¤ºÉ°œÎµ¤µÁ¡µ³Á¨¸Ê¥Š (culture) ¤´„‹³¤„µ¦­¸ ´ŠÁ‡¦µ³®r­Š¨—¨Š ‹¹Š¤¸‡ªµ¤‹ÎµÁž}œ°¥nµŠ¥·ÉŠš¸É‹³˜o°ŠÁ¡¤­µ¦¡ª„‡µ¦·É r°œ‹µ„®¨nŠ £µ¥œ°„¨ŠÅžÄœ°µ®µ¦­Îµ®¦´„µ¦Á¡µ³Á¨¥ŠÁœ¸Ê ºÊ°Á¥ºÉ°¡º Ž¹ÉŠ¤´„Å—o‹µ„œÊ嘵¨Áž}œ­ªœÄ®n n Ž¹ÉŠ

ŗo„n œÊ嘵¨Ž¼Ã‡¦­ (sucrose) „¨¼Ã‡­ (glucose) ¨³¢¦»‡Ã˜­ (fructose) Áž}œ˜oœ

7.1.2.2 ª·˜µ¤·œ (vitamins) ¡º­nªœ¤µ„šÁ¡µ³Á¨¸É ¸Ê¥ŠÄœ­£µ¡ž¨°—ÁºÊ°­µ¤µ¦™­¦oµŠ ª·˜µ¤·œÅ—oÁ°Š ˜n¤¸ž¦·¤µ–ŤnÁ¡¸¥Š¡°‹¹Š˜o°ŠÁ¡·É¤ª·˜µ¤œ¨ŠÄœ°µ®µ¦­· ´ŠÁ‡¦µ³®rÁ¡ºÉ°Ä®o¡ºÁ‹¦·Á˜· Øŗo°¥nµŠž„˜· ÁœºÉ°Š‹µ„ª·˜µ¤·œ¤¸®œoµš¸ÉÁž}œ coenzyme Ĝ ªœ„µ¦˜nµŠÇ œ·—š¸É¤¸„µ¦Äo„´œ¤µ„

Ĝ°µ®µ¦­Îµ®¦´„µ¦Á¡µ³Á¨¥ŠÁœ¸Ê ºÊ°Á¥ºÉ°¡º‡º° Ś°µ¤¸œ (thiamine, B1) 7.1.2.3 „¦—°³¤·Ãœ (amino acid) „¦—°³¤·Ãœš´ÊŠ®¨µ¥Å¤n¤¸‡ªµ¤‹ÎµÁžœ˜} n°„µ¦Á‹¦· Á˜·Ã˜ °ŠÁœ°Á¥ºÊ ºÉ°¡º ¥„ÁªœÅ„¨Žo ¸œ (glycine) Ž¹ÉŠ¤¸„µ¦ÄoĜ®¨µ¥­¼˜¦°µ®µ¦ „µ¦Á˜·¤„¦—°³¤·Ãœ ¨ŠÅžÄœ°µ®µ¦Á¡µ³Á¨¸Ê¥ŠÁœ°Á¥ºÊ ºÉ°œ´ÊœÁ¡ºÉ°Á¡·É¤„µ¦Á‹¦· °ŠÁŽ¨¨r ­nŠÁ­¦·¤„µ¦¡´•œµÅžÁž}œ˜oœ °Š ¡º 7.1.2.4 ­µ¦ž¦³„°›¦¦¤µ˜·š¸ÉŽ´Žo°œ (natural complexes) ­µ¦ž¦³„°š¸ÉŽ´ Žo°œš¸É¤¸„µ¦œµ¤µÄÎ o­nªœÄ®nœ´Êœ ŗo‹µ„­µ¦­„´—‹µ„¡º Ánœ œÊ夳¡¦oµª œÊ夳Á ºÉ°Áš« „¨oª¥®°¤ œÊ娳ÁœºÊ°¤´œ ¦´ÉŠ ­µ¦­„´—‹µ„¥¸­˜r ¨³ protein hydrolysates (Ánœ casine hydrolysate, peptone ¨³ tryptone) 13

œÊε˜o¤¤´œ ¦´ÉŠ Ĝ¤´œ ¦´ÉŠ¤¸Ã¡¨¸Á°¤¸œ (polyamine) ¨³ biosynthetic enzyme „¦³‹µ¥ °¥¼nš´ÉªÅžÄœ­ªœ˜n nµŠÇ Ž¹ÉŠ­µ¦Ã¡¨¸Á°¤¸œ¤¸Ÿ¨˜n°„µ¦Á‹¦·Â¨³¡´•œµ °ŠÁŽ¨¨r ×¥ÁŒ¡µ³¤¸Ÿ¨˜n° „µ¦Á¡·É¤ž¦·¤µ– °Š„¦—œª‡¨· ¸°·‡ (nucleic acid) šÎµÄ®oÁ„·—„µ¦ÂŠÁŽ¨¨n rÅ¤Ã˜Ž·­ (mitosis) Ĝ ÁœºÊ°Á¥ºÉ°¤µ„ ¹Êœ œ°„‹µ„œ¸ÊĜ¤´œ ¦´ÉŠ¥´Šž¦³„°—ª¥Âžo jŠ œÊ嘵¨ Þ¦˜œÂ¨³ª¸ ·˜µ¤·œ°¸„—oª¥ „µ¦Ä­n œÊε˜o¤¤´œ ¦´ÉŠÄœ­¼˜¦°µ®µ¦Á¡µ³Á¨¸Ê¥ŠÁœºÊ°Á¥ºÉ°„¨oª¥Å¤o ¤¸¦µ¥Šµœªnµ­nŠŸ¨Ä®oÁ¤¨È—Š°„Å—o—¸ ¹ÊœÂ¨³˜oœ °n°œ¤¸‡ªµ¤Â Ȋ¦Š (Arditti and Ernst 2008) 7.1.2.5 ŸŠ™nµœ„´¤¤´œ˜ r (activated charcoal) ŸŠ™nµœŸ¨·˜¤µ‹µ„Ťoš¸É™¼„‡µ¦r°ÅœŽr —oª¥°–®£» ¼¤·­¼Š‡ª‡¼nŞ„´Å°œÊε £µ¥ÄœÁž}œ¦¼š¸ÉÁž}œ˜µ nµ¥¨³Á°¸¥— Ž¹ÉŠ­µ¤µ¦™—¼—„p­®¦º°nª¥—¼— Ž´­µ¦¡·¬ °Š­µ¦ž¦³„°¢eœ°¨ ¨³Á¤¨µœ·œ (melanin) šÎµÄ®Á„o ·—­£µ¡—ε¤º— °Š°µ®µ¦­nŠŸ¨ Ä®oÁ„·—¦µ„Å—o—¸ ¨³nª¥‡Š­£µ¡Á­™¸¥¦ °Š¡¸Á°Ž (pH) (‡Îµœ¼œ 2544) 7.2 ­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ °Š¡º (plant growth regulator) ®¦º° ±°¦r䜡ºÁž}œ­µ¦š¸É¡º ­¦oµŠ ¹Êœ ¤oªnµ‹³¤¸Äœž¦¤µ–Á¡· ¥ŠÁ¨¸ Ȅœo°¥„Ȥ¸Ÿ¨˜n°„µ¦Áž¨¸É¥œÂž¨Š­£µ¡šµŠ­¦¸¦ª·š¥µ °Š¡º

×¥¤¸šµš­Îµ‡´Äœ„µ¦„¦³˜»oœ®¦º°¥´¥´ÊŠ„µ¦Á‹¦·Á˜Ã˜Äœ¡· ºœœÇ´Ê Ž¹ÉŠ­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ °Š¡ºš¸Éœ·¥¤œÎµ¤µÄoĜ„µ¦Á¡µ³Á¨¸Ê¥ŠÁœ°Á¥ºÊ ºÉ°¡º ŗ„o n °°„Ž·œ(auxin) ¨³ ŎØҜ·œ (cytokinin) ­nªœ„¨»n¤°ºÉœš¸É‹´—Áž}œ­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ °Š¡º ‡º° gibberellins abscisic acid ¨³ ethylene œ´ÊœÅ¤n‡n°¥œ·¥¤Äo ‹³ÄooµŠÄœ¡µŠœº ·—Ášnµœ´Êœ (»¬¦µ£¦–r 2548)

7.3 ª»oœ (gelling agents) ‹´—Áž}œ¡ª„¡°¨·ÂŽÈ„‡µÅ¦—rš¸É¤¸¤ª¨Ã¤Á¨„»¨­¼Š nª¥šÎµÄ®°µ®µ¦Â o Ȋ

˜´ª ª»oœ­nªœÄ®nš¸ÉčoĜ„µ¦Á¡µ³Á¨¸Ê¥ŠÁœ °Á¥ºÊ ºÉ°Å—o¤µ‹µ„Ÿ¨Ÿ¨·˜ °Š­·ÉŠ¤¸¸ª·˜ ‡º° ­µ®¦nµ¥œÊεÁ‡È¤ ¤¸ ®¨µ¥œ·—ŗ„o n agar agarose ¨³ gelrite (Arditti and Ernst 2008)

8. „µ¦Á¡µ³Á¨¸Ê¥ŠÁœºÊ°Á¥ºÉ°„¨oª¥Å¤o¦°ŠÁšoµœµ¦¸Äœ­£µ¡ž¨°—ÁºÊ° ­Îµ®¦´„µ¦œÎµÁš‡ÃœÃ¨¥¸„µ¦Á¡µ³Á¨¸Ê¥ŠÁœ°Á¥ºÊ ºÉ°¡ºŸ¨·˜„¨oª¥Å¤o¦°ŠÁšoµœµ¦¸ ŗo¤¸„µ¦ ª·‹´¥Â¨³¡•œµ‹µ„®´ o°Šž’·´˜·„µ¦Äœ®¨µ¥ž¦³Áš« Ánœ ‹¸œ Řo®ª´œ ¨³°·œÁ—¸¥ Áž}œ˜oœ [Chen et al. (2004), Sama et al. (2004), Yung and Nean (2001) Huang et al. (2001)] Á¡ºÉ°Ä®oÁ„·—«´„¥£µ¡ Ĝ„µ¦Ÿ¨·˜„¨ª¥Å¤o o¦°ŠÁšoµœµ¦¸Äœž¦·¤µ–¤µ„˜µ¤˜o°Š„µ¦Â¨³­µ¤µ¦™œÎµ°°„ž¨¼„Å—oĜ­£µ¡ ›¦¦¤µ˜· ×¥¤¸¦µ¥Šµœ„µ¦ª·‹´¥Á„¥ª„¸É ´„¨ª¥Å¤o o¦°ŠÁšoµœµ¦¸ ¡°­¦»žÅ—o—´Šœ ¸Ê Lin et al. (2000) «¹„¬µ„µ¦ ¥µ¥¡œ›´ »r„¨oª¥Å¤o Paphiopedilum hybrid (Paphio- pedilum callosum ‘Oakhi’ x Paph. Lawreneanum ‘Tradition’) —oª¥ª·›¸„µ¦Á¡µ³Á¨¥ŠÁœ¸Ê ºÊ°Á¥ºÉ°‹µ„„µ¦ Á¡µ³Á¨¸Ê¥ŠÂ‡¨¨´­š¸É´„œÎµ‹µ„ Protocorm Ž¹ÉŠÅ—o‹µ„„µ¦Á¡µ³Á¤¨È— ¡ªnµ ­¼˜¦°µ®µ¦ 1/2 Murashige and Skoog, 1962 (MS) š¸Éž¦³„°—ª¥­µ¦‡ª‡o »¤„µ¦Á˜·Ã˜ Dichlorophenoxyacetic acid (2,4-D) 14

‡ªµ¤Á o¤ oœ 1-10 ¤·¨¨·„¦´¤˜n°¨·˜¦ ¨³ Thidiazuron (TDZ) ‡ªµ¤Á o¤ oœ 0.1-1 ¤·¨¨·„¦´¤˜n°¨·˜¦ ‡¨¨´­¤¸„µ¦Á‹¦·—¸Á¤ºÉ°¥oµ¥ÅžÁ¡µ³Á¨¸Ê¥Šœ­¼˜¦°µ®µ¦Á—·¤ ˜n‹³¤¸ž¦¤µ–Á¡· ·É¤¤µ„ ¹ÊœÁ¤ºÉ°Á¡µ³Á¨¸Ê¥Š œ­¼˜¦°µ®µ¦ 1/2 MS š¸Éž¦³„°—ª¥­µ¦‡ª‡o »¤„µ¦Á˜·Ã˜ 2,4-D ‡ªµ¤Á o¤ oœ 5 ¤·¨¨·„¦´¤˜n° ¨·˜¦ ¨³ TDZ ‡ªµ¤Á o¤ oœ 1 ¤·¨¨·„¦´¤˜n°¨·˜¦ Ž¹ÉŠ­µ¤µ¦™¡´•œµÁž}œ protocorm-like bodies (PLBs) ¨³˜oœš¸É­¤¼¦–r ­µ¤µ¦™¥oµ¥¨Šž¨¼„Äœ„¦³™µŠÂ¨³¤¸„µ¦Á‹¦·Á˜·Ã˜š¸É—¸ Huang et al. (2001) «¹„¬µ„µ¦ ¥µ¥¡œ›´ »r„¨oª¥Å¤o Paphiopedilum hybrid —oª¥ª·›¸„µ¦ Á¡µ³Á¨¸Ê¥ŠÁœºÊ°Á¥ºÉ° ×¥—´—ž¨Š­nªœž¦³„° °Š°µ®µ¦Á¡ºÉ°´„œÎµÄ®Á„o ·—¥°—°¥nµŠ¦ª—Á¦Èª ץčo ­¼˜¦°µ®µ¦ MS ¦nª¤„´ Murashige & Tucker, 1969 (MT) ª·˜µ¤·œ, ń¨Ž¸œ ¨³°·œÃœŽ·š°¨ ¦nª¤„´­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ 6-benzyladenine (BA) ‡ªµ¤Á o¤ oœ 13 ŤÇ¦Ã¤¨µ¦r Į- Naphthalene acetic acid (NAA) ‡ªµ¤Á o¤ oœ 1.6 ŤÇ¦Ã¤¨µ¦r adenine sulfate (Ads) 0.15 ¤·¨¨·Ã¤

¨µ¦r NaH2PO4.H2O 1.23 ¤·¨¨·Ã¤¨µ¦r œÊ嘵¨ 0.18 ä¨ ¨³œÊ夳¡¦oµª‹µ„Ÿ¨¤³¡¦oµª°n°œ 15 Áž°¦rÁŽÈœ˜r (v/v) ®¦º° casein hydrolysate 1 „¦´¤˜n°¨·˜¦ ®¦º°œÊε˜o¤¤´œ ¦´ÉŠ (potato homogenate) 10

„¦´¤˜n°¨·˜¦ Á¡ºÉ°Á¡·É¤‹Îµœªœ¥°—šª¸‡¼–¨³„µ¦´„œÎµÄ®oÁ„—¦µ„· Á¡¸¥Š œ˜°œÁ—´Ê ¸¥ª®¦º°œ ­¼˜¦°µ®µ¦Á—¸¥ª‹µ„¥°—š¸Éž¨°—ÁºÊ° °Š˜oœ„¨oµ‹µ„Ÿ¼ož¦ž¦´ »Š¡´œ›»rÁ¡°„µ¦‡ºÉ oµ ¡ªnµ TDZ ­nŠŸ¨ ¥´¥´ÊŠ„µ¦Á¡·É¤‹Îµœªœ °Š¥°—¨³¦µ„ œÊ嘵¨¤°¨Ã˜­ ¨—„µ¦Á„·—¦µ„ Ž¹ÉŠª·›¸„µ¦ oµŠ˜oœ­µ¤µ¦™Á¡·É¤ ‹Îµœªœ °Š Paphiopedilum ŗoš»„Ç 12 ­´ž—µ® r

Yung and Nean (2001) «¹„¬µ°µ¥» °Š {„š¸ÉÁ®¤µ³­¤˜n°„µ¦Š°„ °ŠÁ¤¨È— ­¼˜¦°µ®µ¦

1/2 MS, 1/8 MS ¨³ Thomale GD media, 1954 (TM) Ĝ„µ¦Á¡µ³Á¨¸Ê¥ŠÁ¤¨— °Š„¨È oª¥Å¤o Paphiopedilum primulinum ×¥š—¨°ŠÄ­n œÊ夳¡¦oµª 100 ¤·¨¨·¨·˜¦˜n°¨·˜¦ œÊε¤´œ ¦´ÉŠ˜o¤ 20 „¦´¤˜n°¨·˜¦ š¦·žÃ˜œ 2 „¦´¤˜n°¨·˜¦ ¨³ œÊε˜o¤„¨oª¥ 20 „¦´¤˜n°¨·˜¦ ¡ªnµ °µ¥» {„š¸ÉÁ®¤µ³­¤ ˜n°„µ¦Š°„ °ŠÁ¤¨È—‡º° 110 ª´œ ®¨´Š‹µ„„µ¦Ÿ­¤Á„­¦ ¨³¡ªnµ „µ¦Ä­nœÊ夳¡¦oµª­µ¤µ¦™ „¦³˜»oœ„µ¦Š°„ °ŠÁ¤¨È—Å—o—¸š¸É­»— ¦°Š¨Š¤µ‡º° œÊε¤´œ ¦´ÉŠ˜o¤ ¨³š¦·žÃ˜œ ˜µ¤¨Îµ—´ ­nªœœÊε˜o¤ „¨oª¥Ä®oŸ¨¥¥´ ´ÊŠ„µ¦Š°„ °ŠÁ¤¨È— œ°„‹µ„œ¸Ê¡ªnµ ŸŠ™nµœ„´¤¤´œ˜r¤¸Ÿ¨˜n°‡ªµ¤­µ¤µ¦™Äœ„µ¦ Š°„ °ŠÁ¤¨È—œo°¥ ˜n­nŠŸ¨˜n°„µ¦¡´•œµ °Š protocorm ¨³„µ¦Á¨¸Ê¥ŠÄœ°µ®µ¦Á®¨ª¤¸°´˜¦µ„µ¦ Š°„ °ŠÁ¤¨È—­¼Š„ªnµ„µ¦Á¨¸Ê¥Šœ°µ®µ¦Â Ȋ Chen et al. (2002) «¹„¬µ„µ¦Á„·—¥°—šª¸‡¼–‹µ„˜µ °Š„oµœn°—°„„¨oª¥Å¤o Paphio- pedilum philippinense 2 ­µ¥¡´œ›»r (PH59 ¨³ PH60) ×¥Á¡µ³Á¨¸Ê¥Šœ­¼˜¦°µ®µ¦ 1/2 MS š¸ÉŤn¤¸ ­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ ¨³ž¦³„°—ª¥­µ¦‡ª‡o »¤„µ¦Á˜·Ã˜ 2,4-D ¨³ TDZ Áž}œÁª¨µ 6 Á—º°œ ¡ªnµ ­¼˜¦°µ®µ¦š¸ÉŤn¤¸­µ¦‡ª‡»¤„µ¦Á˜Ã˜¤· ¸·ÊœÁœºÊ°Á¥ºÉ°š¸ÉÁ„·—¥°— 33.3 ¨³ 0 Áž°¦rÁŽÈœ˜r ¨³¤¸ ‹Îµœªœ¥°—˜°n ·ÊœÁœºÊ°Á¥ºÉ° 1 ¨³ 0 ·Êœ ‹µ„­µ¥¡´œ›»r PH59 ¨³ PH60 ˜µ¤¨Îµ—´ Ĝ­µ¥¡´œ›»r PH59 15

¡ªnµ­¼˜¦°µ®µ¦š¸Éž¦³„°—oª¥­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ 2,4-D ‡ªµ¤Á o¤ oœ 4.52 ŤÇ¦Ã¤¨µ¦r ¦nª¤„´ TDZ ‡ªµ¤Á o¤ oœ 0.45 ŤÇ¦Ã¤¨µ¦r ¤¸·ÊœÁœ°Á¥ºÊ ºÉ°š¸ÉÁ„—¥°—¨³‹· 圪œ¥°—˜n°·ÊœÁœºÊ°Á¥ºÉ° ¤µ„š¸É­»— 66.7 ¨³ 1.5 Áž°¦rÁŽÈœ˜r ˜µ¤¨Îµ—´ ­nªœÄœ­µ¥¡œ›´ »r PH60 ¡ªnµ ­¼˜¦°µ®µ¦š¸É ž¦³„°—ª¥­µ¦‡ª‡o »¤„µ¦Á˜·Ã˜ 2,4-D ‡ªµ¤Á o¤ oœ 4.52 ŤÇ¦Ã¤¨µ¦r ¦nª¤„´ TDZ ‡ªµ¤ Á o¤ oœ 0.45 ŤÇ¦Ã¤¨µ¦r ¤¸·ÊœÁœºÊ°Á¥°šºÉ ¸ÉÁ„·—¥°—¤µ„š¸É­»— 80 Áž°¦rÁŽÈœ˜r Ž¹ÉŠÄœ­¼˜¦°µ®µ¦š¸É ž¦³„°—ª¥­µ¦‡ª‡o »¤„µ¦Á˜·Ã˜ 2,4-D ‡ªµ¤Á o¤ oœ 4.52 ŤÇ¦Ã¤¨µ¦r Á¡¸¥Š°¥nµŠÁ—¥ª¸ ­µ¤µ¦™´„œÎµÄ®o¤¸‹Îµœªœ¥°—˜n°·ÊœÁœºÊ°Á¥ºÉ°¤µ„š¸É­»— 7.0 ¥°— ¨³¥°—š¸Éŗo­µ¤µ¦™¡´•œµÁž}œ˜oœ š¸É­¤¼¦–rŗoÁ¤ºÉ°¥oµ¥ÅžÁ¨¸Ê¥Šœ­¼˜¦°µ®µ¦š¸ÉŤn¤¸­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ Áž}œÁª¨µ 3 Á—º°œ Lee and Lee (2003) «¹„¬µ„µ¦ ¥µ¥¡´œ›»r„¨oª¥Å¤o Cypripedium formosanum ×¥„µ¦ ´„œÎµÄ®oÁ„—‡¨¨· ´­‹µ„ protocorm š¸Éŗo‹µ„„µ¦Á¡µ³Á¤¨È— ¤µÁ¡µ³Á¨¸Ê¥Šœ­¼˜¦°µ®µ¦ 1/4 MS š¸É ž¦³„°—ª¥­µ¦‡ª‡o »¤„µ¦Á˜·Ã˜ 2,4-D ‡ªµ¤Á o¤ oœ 4.52 ŤÇ¦Ã¤¨µ¦r ¨³ TDZ ‡ªµ¤Á o¤ oœ 4.54 ŤÇ¦Ã¤¨µ¦r ¡ªnµ ‡¨¨´­­µ¤µ¦™Á¡·É¤ž¦·¤µ–—Á¤¸ ºÉ°¥oµ¥ÅžÁ¡µ³Á¨¸Ê¥Šœ­¼˜¦°µ®µ¦Á—·¤ ¨³

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Chen et al. (2004b) «¹„¬µ„µ¦ ¥µ¥¡ œ›´ »r„¨oª¥Å¤o Paphiopedilum hybrid 2 ­µ¥¡œ›´ »r (PH59 ¨³ PH60) —oª¥ª·›¸„µ¦Á¡µ³Á¨¸Ê¥ŠÁœºÊ°Á¥ºÉ° ×¥„µ¦´„œÎµÄ®oÁ„·—¥°—‹µ„¦Áª–µ—Ÿ¨ °Š· ·Êœ­nªœÄ Á¤°Á¡µ³Á¨ºÉ ¸Ê¥Šœ­¼˜¦°µ®µ¦ MS š¸É—´—ž¨Š›µ˜»°µ®µ¦ (1/2 strength macro-and full- strength micro elements) Ž¹ÉŠÅ¤n¤¸­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ Ĝš¸É¤º—Áž}œ¦³¥³Áª¨µ 1 Á—º°œ ­nªœŸ­¤ °Š­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ 2,4-D (0, 4.52 ¨³ 45.25 ŤÇ¦Ã¤¨µ¦r) ¨³ TDZ (0, 0.45, 4.54 ¨³ 22.71 ŤÇ¦Ã¤¨µ¦r) čoš—­°„µ¦´„œÎµÄ®oÁ„·—¥°— °Š·Êœ­nªœÄ ¡ªnµ Ĝ­µ¥¡´œ›»r PH59 ­¼˜¦°µ®µ¦š¸Éž¦³„°—ª¥­µ¦‡ª‡o »¤„µ¦Á˜·Ã˜ TDZ ‡ªµ¤Á o¤ oœ 4.54 ŤÇ¦Ã¤¨µ¦r ­µ¤µ¦™ Á¡·É¤‹Îµœªœ¥°—ÁŒ¨¸É¥˜n°·Êœ­ªœÄšn č¸É oš—¨°ŠÅ—o—¸š¸É­»— Ĝ­µ¥¡œ›´ »r PH 60 ­¼˜¦°µ®µ¦š¸É ž¦³„°—ª¥­µ¦‡ª‡o »¤„µ¦Á˜·Ã˜ 2,4-D ‡ªµ¤Á o¤ oœ 4.52 ŤÇ¦Ã¤¨µ¦r ¦nª¤„´ TDZ 0.45 Ť Ǧ䨵¦r ­µ¤µ¦™´„œÎµÄ®oÁ„·—¥°—‹µ„·Êœ­nªœÄšÄ¸É oš—¨°ŠÅ—o—¸š¸É­»— œ°„‹µ„œ¸Ê ­¼˜¦°µ®µ¦š¸É ž¦³„°—ª¥­µ¦‡ª‡o »¤„µ¦Á˜·Ã˜ 2,4-D ‡ªµ¤Á o¤ oœ 4.52 ŤÇ¦Ã¤¨µ¦r TDZ 4.52 ŤÇ¦Ã¤ ¨µ¦r ¨³ 2,4-D ‡ªµ¤Á o¤ oœ 4.52 ŤÇ¦Ã¤¨µ¦r ¦nª¤„´ TDZ ‡ªµ¤Á o¤ oœ 4.54 ŤÇ¦Ã¤¨µ¦r ´„ œÎµÄ®oÁ„·—‹Îµœªœ¥°—ÁŒ¨¸É¥˜°n ·Êœ­nªœÄ¤µ„„ªnµ­¼˜¦°µ®µ¦š¸ÉŤn¤¸­µ¦‡ª‡»¤„µ¦Á˜Ã˜· ¡ºš¸É 16

­¤¼¦–r‹³¤¸‹Îµœªœ¦µ„ÁŒ¨¸É¥ 1-3 ¦µ„ š¸ÉÁ„·—‹µ„„µ¦¥oµ¥¥°—šÅ—¸É oŞÁ¨¸Ê¥Šœ­¼˜¦°µ®µ¦š¸ÉŤn¤¸­µ¦ ‡ª‡»¤„µ¦Á˜Ã˜Áž· }œÁª¨µ 22 Á—º°œ Sama and Kalita (2004) «¹„¬µŸ¨„µ¦˜°­œ°Š °ŠÁ¤¨È— Paphiopedilum insigne Ĝ

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¤·¨¨·Á¤˜¦ ‹Îµœªœ 1 PLBs ĜÁª¨µ 1 že Lin et al. (2004) «¹„¬µ„µ¦Š°„ °ŠÁ¤¨È—‹µ„ {„ °Š Paphiopedilum armeniacum ¨³ Paphiopedilum micranthum °µ¥» 120 ª´œ ¨³ 180 ª´œ ®¨´Š‹µ„„µ¦Ÿ­¤Á„­¦ Á¤ºÉ°œÎµÅžÁ¡µ³Á¨¥Š¸Ê œ­¼˜¦°µ®µ¦ 1/4 MS, MS, Rappaport, Konforti and Navon, 1956 (R), KC, VW ¨³ Hyponex 1 ×¥Á„ȝŸ¨®¨Š‹µ„Á¨´ ¸Ê¥ŠÄœ­˜¦°µ®µ¦˜¼ nµŠÇ Áž}œÁª¨µ 120 ª´œ ¡ªnµ Ĝ­¼˜¦°µ®µ¦š¸É¤¸Â¦n›µ˜»œo°¥¤¸ °´˜¦µ„µ¦Š°„ °ŠÁ¤¨È— ¤µ„„ªnµÄœ­¼˜¦°µ®µ¦š¸É¤¸Â¦n›µ˜»¤µ„ „µ¦Š°„ °ŠÁ¤¨È—¤¸¤µ„š¸É­»—Äœ­¼˜¦ °µ®µ¦ R ¨³œÊ夳¡¦oµª¤¸­nªœª¥„¦³˜n »oœÄ®oÁ„—„µ¦Š°„ °ŠÁ¤¨· ȗ „µ¦Á˜·¤ŸŠ™nµœ „´¤¤´œ˜r ž¦·¤µ– 2 „¦´¤˜n°¨·˜¦ ¨³„¨oª¥— ž¦·¤µ– 100 „¦´¤˜n°¨·˜¦ šÎµÄ®o„µ¦Á‹¦·Á˜·Ã˜Â¨³¡´•œµ °Š ˜oœ„¨oµ—¸ ¹Êœ Yan et al. (2006) «¹„¬µ„µ¦ ¥µ¥¡´œ›»r„¨oª¥Å¤o Cypripedium flavam —oª¥ª·›¸„µ¦Á¡µ³ Á¨¸Ê¥ŠÁœºÊ°Á¥ºÉ° ×¥„µ¦´„œÎµÄ®oÁ„·—¥°—šª¸‡¼–‹µ„˜œ°o n°œš¸Éŗo‹µ„„µ¦Á¡µ³Á¤¨È—¤µÁ¡µ³Á¨¸Ê¥Šœ­¼˜¦ °µ®µ¦ Harvais, 1974 (Harvais) š¸Éž¦³„°—oª¥­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ÅŽÃ˜Å‡œ·œ 2 œ·— (N- Benzyl-9-(2-tetrahydropyranyl) adenine (BAP) ®¦º° Kn) ¦nª¤„´œµ˜ÊÎ o¤¤´œ ¦´ÉŠ š¸É¤¸‡ªµ¤Á o¤ oœ 17

˜„˜nµŠ„´œ ¡ªnµ ­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ BAP ­µ¤µ¦™´„œÎµÄ®oÁ„—¥°—šª· ¸‡¼–¤µ„„ªnµ­µ¦‡ª‡»¤ „µ¦Á˜·Ã˜ Kn ­¼˜¦°µ®µ¦ Harvais š¸Éž¦³„°—ª¥­µ¦‡ª‡o »¤„µ¦Á˜Ã˜· BAP ‡ªµ¤Á o¤ oœ 2.22 ŤÇ¦Ã¤¨µ¦r ¦nª¤„´œÊ嘤¤o ´œ ¦´ÉŠ ž¦·¤µ– 20 „¦´¤˜n°¨·˜¦ ­µ¤µ¦™´„œÎµÄ®oÁ„—¥°—šª· ¸‡¼–¤µ„ š¸É­»— 95 Áž°¦rÁŽÈœ˜r °Š˜oœ°n°œš¸Éčoš—¨°Š ¨³¤¸¥°—Á„·—¤µ„š¸É­»— 2.55 ¥°—˜n°˜oœ°n°œ 1 ˜oœ ­Îµ®¦´„µ¦´„œÎµÄ®oÁ„·—¦µ„¨³„µ¦¦°—¸ª·˜ ­¼˜¦°µ®µ¦ 1/2 Harvais š¸É¤¸ŸŠ™nµœ„´¤¤´œ˜rž¦·¤µ– 0.6 „¦´¤˜n°¨·˜¦ ­nŠŸ¨˜n°„µ¦Á„·—¦µ„¨³¤¸„µ¦¦°—¸ª·˜­¼Šš¸É­»— ‹µ„„µ¦«¹„¬µœ¸Ê­—Šªnµ„¨oª¥Å¤o Cypripedium flavam ­µ¤µ¦™ ¥µ¥¡´œ›»r×¥„µ¦šÎµÄ®oÁ„—¥°—šª· ¸‡¼–‹µ„˜oœ°n°œš¸Éŗo‹µ„„µ¦Á¡µ³ Á¤¨È— Hong et al. (2008b) «¹„¬µ„µ¦ ¥µ¥¡´œ›»r„¨oª¥Å¤o Paphiopedilum ‘Alma Gavaert’ ‹µ„ PLBs ×¥˜¦ŠÂ¨³„µ¦Á„·—¥°—šª¸‡¼–‹µ„‡¨¨´­š¸É¡´•œµ‹µ„Á¤¨È— ×¥´„œÎµÁ¤¨È—š¸Éŗo‹µ„ „°µ¥{ » 5 Á—º°œ Ä®o¡´•œµÁž}œÂ‡¨¨´­œ­¼˜¦°µ®µ¦ 1/2 MS š¸Éž¦³„°—ª¥­µ¦‡ª‡o »¤„µ¦Á˜·Ã˜ 2,4-D ‡ªµ¤Á o¤ oœ 22.6 ŤÇ¦Ã¤¨µ¦r ¨³ TDZ ‡ªµ¤Á o¤ oœ 4.54 ŤÇ¦Ã¤¨µ¦r Ĝš¸É¤º— ‡¨¨´­¤¸„µ¦

Á¡·É¤ž¦·¤µ–°¥µŠ¤µ„n ¨³Å¤n¤¸„µ¦Áž¨¸É¥œÂž¨Š¨´„¬–³šµŠ­´–“µœªš¥µÁ¤· ºÉ°¥oµ¥¨Šœ­¼˜¦°µ®µ¦ Á—·¤š»„Ç 2 Á—º°œ Áž}œÁª¨µ¤µ„„ªnµ 2 že ¨³Á¤ºÉ°¥oµ¥Â‡¨¨´­ÅžÁ¡µ³Á¨¸Ê¥Šœ­¼˜¦°µ®µ¦ 1/2 MS š¸É ž¦³„°—ª¥­µ¦‡ª‡o »¤„µ¦Á˜·Ã˜ NAA ‡ªµ¤Á o¤ oœ 26.85 ŤÇ¦Ã¤¨µ¦r ‡¨¨´­¡´•œµÁž}œ PLBs ®¦º° ¥°— ÁŒ¨¸É¥ 4.7 PLBs ˜n°¥°— ‹µ„‡¨¨´­Â˜n¨³·Êœš¸ÉšÎµ„µ¦š—¨°Š®¨´Š‹µ„Á¡µ³Á¨¸Ê¥ŠÁž}œ

Áª¨µ 120 ª´œ ®¨´Š‹µ„œ´Êœ 72 ¨³ 240 ª´œ °Š„µ¦Á¡µ³Á¨¸Ê¥Šœ­¼˜¦°µ®µ¦Á—·¤ šÎµÄ®oŗo˜µ¥°—

‹Îµœªœ 25 ˜µ¥°—¨³¡ºš¸É­¤¼¦– r‹Îµœªœ 75 ˜oœ ‹µ„„µ¦´„œÎµÄ®Á„o —¥°—šª· ¸‡¼–‹µ„„µ¦ Á¡µ³Á¨¸Ê¥ŠÁ¦·É¤˜oœ ­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ Kn ‡ªµ¤Á o¤ oœ 4.65 ŤÇ¦Ã¤¨µ¦r ¤¸‡ªµ¤Á®¤µ³­¤ ­Îµ®¦´„µ¦´„œÎµÄ®oÁ„·—¥°—šª¸‡¼–¨³­µ¤µ¦™´„œÎµÄ®Á„o ·—¥°—ÁŒ¨¸É¥ 3 ¥°— ®¨´Š‹µ„Á¡µ³Á¨¸Ê¥Š ¥°—°n°œÁžœÁª¨µ} 60 ª´œ ¡ºš¸Éŗo‹µ„„µ¦Á¡µ³Á¨¸Ê¥Š¤„µ¦Á‹¦¸ ·Á˜·Ã˜Áž}œž„˜·Á¤ºÉ°¥µ¥Åžž¨o ¼„—oª¥ ­Â¢„œ´É¤¤°­ÄœÁ¦º°œ„¦³‹„ Ng and Saleh (2010a) «¹„¬µ„µ¦ ¥µ¥¡œ›´ »r„¨oª¥Å¤o Paphiopedilum rothschildianum ‹µ„ primary PLBs Ä®oÁ„·— secondary PLBs —oª¥ª·›¸„µ¦Á¡µ³Á¨¸Ê¥ŠÁœ°Á¥ºÊ ºÉ° ×¥Á¡µ³ Á¨¸Ê¥Šœ­¼˜¦ °µ®µ¦ 1/2 MS š¸Éž¦³„°—oª¥­µ¦‡ª‡µ„µ¦Á‹¦·Á˜Ã˜· BA ‡ªµ¤Á o¤ oœ 1.0, 2.0, 3.0 ¨³ 4.0 ŤÇ¦Ã¤¨µ¦r ®¦º° Kn ‡ªµ¤Á o¤ oœ 1.0, 2.0, 3.0 ¨³ 4.0 ŤÇ¦Ã¤¨µ¦r ¡ªnµ ­¼˜¦°µ®µ¦š¸É ­µ¤µ¦™´„œÎµÄ®oÁ„·— secondary PLBs ŗo—¸š¸É­»—‡º° ­¼˜¦°µ®µ¦ 1/2 MS š¸Éž¦³„°—oª¥­µ¦‡ª‡»¤ „µ¦Á˜·Ã˜ Kn ‡ªµ¤Á o¤ oœ 4.0 ŤÇ¦Ã¤¨µ¦r ×¥¤¸‹Îµœªœ secondary PLBs Á„·— ¹ÊœÁŒ¨¸É¥ 4.1 PLBs ˜n° 1 primary PLBs Ĝ¦³¥³Áª¨µ 8 ­´ž—µ®r ¨³Á¤ºÉ°Á¡µ³Á¨¸Ê¥Š secondary PLBs š¸ÉÁ„·— ¹Êœœ 18

­¼˜¦°µ®µ¦ 1/2 MS š¸ÉŤn¤¸­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ ˜nž¦³„°—oª¥œÊ嘤„¨o oª¥‡ªµ¤Á o¤ oœ 60 „¦´¤ ˜n°¨·˜¦ ­µ¤µ¦™´„œÎµÄ®oÁ„·— secondary PLBs Ä®¤n ÁŒ¨¸É¥ 9.5-12.1 PLBs. Ng and Saleh (2010b) «¹„¬µ„µ¦ ¥µ¥¡œ›´ »r„¨oª¥Å¤o Paphiopedilum rothschildianum —oª¥ª·›¸„µ¦Á¡µ³Á¨¸Ê¥ŠÁœºÊ°Á¥ºÉ° ×¥„µ¦´„œÎµÄ®oÁ„—¥°—šª· ¸‡¼–‹µ„˜µ oµŠÂ¨³¥°—×¥Á¡µ³Á¨¸Ê¥Šœ ­¼˜¦°µ®µ¦ 1/2 MS š¸Éž¦³„°—oª¥ organic nitrogen ‡º° casein hydrolysate peptone ¨³ trytone- peptone š¸É‡ªµ¤Á o¤ oœ 0.5, 1.0 ¨³ 2.0 „¦´¤˜n°¨·˜¦ ¡ªnµ ­¼˜¦°µ®µ¦ 1/2 MS š¸Éž¦³„°—ª¥o organic nitrogen ­µ¤µ¦™Á¡·É¤‹Îµœªœ¥°—šª¸‡¼–Å—o¤µ„„ªnµ­¼˜¦°µ®µ¦š¸ÉŤn¤¸ organic nitrogen ‹µ„ š´ÊŠ˜µ oµŠÂ¨³¥°— ×¥¤¸‹Îµœªœ¥°—ÁŒ¨¸É¥ 2.9 ¥°—˜n°˜µ oµŠ ¨³ 2.8 ¥°—˜n°¥°—Á—·¤ ®¨´Š‹µ„ Á¡µ³Á¨¸Ê¥ŠÁž}œÁª¨µ 16 ­´ž—µ®r œ­¼˜¦°µ®µ¦ 1/2 MS š¸Éž¦³„°—ª¥o peptone ‡ªµ¤Á o¤ oœ 1.0 „¦´¤˜n°¨·˜¦ ¨³ tryptone-peptone ‡ªµ¤Á o¤ oœ 2.0 „¦´¤˜n°¨·˜¦ ˜µ¤¨Îµ—´ ¨³Á¤ºÉ°œÎµ¥°—¤µ Á¡µ³Á¨¸Ê¥Šœ°µ®µ¦­¼˜¦Á—·¤Áž}œ¦³¥³Áª¨µ 12 ­´ž—µ®r ‹³Á„—¦µ„· 3-4 ¦µ„ ¨³Á¤ºÉ°¥µ¥¨Šž¨o ¼„Äœ Á¦º°œ„¦³‹„¤¸°´˜¦µ„µ¦¦°—¸ª·˜ 90 Áž°¦rÁŽÈœ˜r

Long et al. (2010) «¹„¬µ°µ¥» °Š {„ œ·— °Š°µ®µ¦ ®¨nŠ °Š‡µ¦r°œ ¨³­µ¦ °·œš¦¸¥r š¸É­nŠŸ¨˜n°„µ¦Š°„ °ŠÁ¤¨È— „µ¦¡´•œµ °ŠÃžÃ˜¦‡°¤ ¨³„µ¦Á‹¦·Á˜·Ã˜ °Š„¨oª¥Å¤ o Paphiopedilum villosum var. densissimum ¡ªnµ ¤¸°´˜¦µ„µ¦ ¥µ¥¡´œ›»rÁ¡·É¤ ¹ÊœÁ¤°ÄºÉ o {„š¸É¤¸°µ¥» 200 ª´œ®¨Š‹µ„Á¡µ³Á¤¨´ ȗ Á¡µ³Á¨¸Ê¥Šœ­¼˜¦°µ®µ¦ KC š¸Éž¦³„°—oª¥œÊ嘵¨„¨¼Ã‡­Â¨³œÊε

¤³¡¦oµª ¨³«¹„¬µŸ¨ °Š­µ¦‡ª‡»¤„µ¦Á˜·Ã˜˜n°„µ¦Á„·—¥°— °Š„¨oª¥Å¤o Paphiopedilum 4

œ·— ¡ªnµ ­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ BA ‡ªµ¤Á o¤ oœ 0.5 ¤·¨¨·„¦´¤˜n°¨·˜¦ ¦nª¤„´ NAA ‡ªµ¤ Á o¤ oœ 0.1 ¤·¨¨·„¦´¤˜n°¨·˜¦ ¨³ BA ‡ªµ¤Á o¤ oœ 0.2 ¤·¨¨·„¦´¤˜n°¨·˜¦ ¦nª¤„´ NAA ‡ªµ¤Á o¤ oœ 0.1 ¤·¨¨·„¦´¤˜n°¨·˜¦ ­µ¤µ¦™Á¡·É¤°´˜¦µ„µ¦Á„·—¥°— °Š„¨oª¥Å¤ o Paphiopedilum villosum var. densissimum ¨³ Paphiopedilum insigne (Lindl.) ˜µ¤¨Îµ—´ ­nªœ­µ¦‡ª‡»¤„µ¦Á˜Ã˜· BA ‡ªµ¤ Á o¤ oœ 5.5 ¤·¨¨·„¦´¤˜n°¨·˜¦ ¦nª¤„´ NAA ‡ªµ¤Á o¤ oœ 0.5 ¤·¨¨·„¦´¤˜n°¨·˜¦ ¨³ BA ‡ªµ¤Á o¤ oœ 1 ¤·¨¨·„¦´¤˜n°¨·˜¦ ¦nª¤„´ NAA ‡ªµ¤Á ¤ o oœ 0.1 ¤·¨¨·„¦´¤˜n°¨·˜¦ ­µ¤µ¦™Á¡¤°·É ´˜¦µ„µ¦Á„·—¥°— °Š„¨oª¥Å¤o Paphiopedilum bellatulum (Rchb. f.) Stein ¨³ Paphiopedilum armeniacum S. C. Chen et F. Y. Liu ŗo¤µ„š¸É­»— ˜µ¤¨Îµ—´

9. „µ¦Á¡µ³Á¨¸Ê¥ŠÁœºÊ°Á¥ºÉ°„¨oª¥Å¤o‹µ„·Êœ­nªœÄÄœ­£µ¡ž¨°—ÁºÊ° Šµœª·‹´¥„µ¦Á¡µ³Á¨¸Ê¥ŠÁœºÊ°Á¥ºÉ°„¨oª¥Å¤o‹µ„œ­·Ê nªœÄ Ážœª} ·›¸„µ¦®œ¹ÉŠš¸Éœ´„ª·‹´¥Á¨º°„čo Ĝ„µ¦ ¥µ¥¡œ›´ »r„¨oª¥Å¤o ÁœºÉ°Š‹µ„Ážœ} œ­·Ê nªœ¡ºš®µŠ¸É nµ¥ ´Êœ˜°œ„µ¦ž’·´˜·Å¤nŽ´Žo°œ ¨³ ­µ¤µ¦™ ¥µ¥¡´œ›»r„¨oª¥Å¤Å—o oÁž}œ‹Îµœªœ¤µ„ ¡°­¦»žÅ—o—´Šœ ¸Ê 19

Nayak et al. (1997) «¹„¬µ„µ¦ ¥µ¥¡œ›´ »r„¨oª¥Å¤o Acampe praemora (Roxb.) Blatter and McCann —oª¥ª·›¸„µ¦Á¡µ³Á¨¸Ê¥ŠÁœºÊ°Á¥ºÉ°‹µ„·Êœ­nªœÄ Á¡ºÉ°´„œÎµÄ®oÁ„·—¥°—×¥˜¦Š ¡ªnµ ­¼˜¦ °µ®µ¦ MS š¸Éž¦³„°—ª¥­µ¦‡ª‡o »¤„µ¦Á˜·Ã˜ TDZ ‡ªµ¤Á o¤ oœ 1 ¤·¨¨·„¦´¤˜n°¨·˜¦ ¤¸„µ¦Á„·— ¥°—¨³‹Îµœªœ¥°—˜n°œ­·Ê nªœ °ŠÄ¤µ„š¸É­»— ¨³¡ªnµ ­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ NAA ‡ªµ¤ Á o¤ oœ˜Éε Ťn­nŠŸ¨Ä®o¤¸„µ¦Á„·—¥°—¤µ„ œÁ¤¹Ê ºÉ°Äo¦nª¤„­µ¦‡ª‡´ »¤„µ¦Á˜·Ã˜ BA Kn ¨³ TDZ °¸„š´ÊŠ­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ NAA ‡ªµ¤Á o¤ oœ­¼Š­nŠŸ¨¥´¥´ÊŠ„µ¦Á„·—¥°— ¥°—š¸ÉÁ„·—‹µ„„µ¦ Á¡µ³Á¨¸Ê¥Š­µ¤µ¦™´„œÎµÄ®oÁ„·—¦µ„Å—o‹µ„­¼˜¦°µ®µ¦ MS š¸Éž¦³„°—oª¥­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ IBA ‡ªµ¤Á o¤ oœ 1 ¤·¨¨·„¦´¤˜n°¨·˜¦ Chen et al. (1999) «¹„¬µ„µ¦ ¥µ¥¡´œ›»r„¨oª¥Å¤ o Oncidium Gower Ramsey —oª¥ª·›¸„µ¦ Á¡µ³Á¨¸Ê¥ŠÁœºÊ°Á¥ºÉ°‹µ„œ­·Ê nªœÄ °ŠÄ°°œn Á¡ºÉ°´„œÎµÄ®oÁ„·— somatic embryos ×¥˜¦Š‹µ„ÁœºÊ°Á¥ºÉ° epidermal ¨³ mesophyll cell š¸É¦·Áª–ž¨µ¥ÄÂ¨³¦Áª–µ—Ÿ¨· ¡ªnµ ­¼˜¦°µ®µ¦ 1/2 MS š¸É ž¦³„°—ª¥­µ¦‡ª‡o »¤„µ¦Á˜·Ã˜ TDZ ‡ªµ¤Á o¤ oœ 0.3-1.0 ¤·¨¨·„¦´¤˜n°¨·˜¦ ­µ¤µ¦™´„œÎµÄ®o

Á„·— somatic embryos £µ¥ÄœÁª¨µ 1 Á—º°œ Chen and Chang (2001) «¹„¬µŸ¨ °Š­µ¦‡ª‡»¤„µ¦Á˜·Ã˜°°„Ž·œ ¨³ÅŽÃ˜Å‡œ·œ ˜n°„µ¦´„œÎµÄ®oÁ„·— somatic embryos ×¥˜¦Š ‹µ„œ­·Ê nªœÄ °Š„¨oª¥Å¤ o Oncidium Gower Ramsey —oª¥ª·›¸„µ¦Á¡µ³Á¨¸Ê¥ŠÁœºÊ°Á¥ºÉ° ¡ªnµ ­¼˜¦°µ®µ¦ 1/2 MS š¸Éž¦³„°—ª¥­µ¦‡ª‡o »¤„µ¦

Á˜·Ã˜°°„Ž·œ (Indole-3-acetic acid (IAA), IBA, NAA ¨³ 2,4-D) ­nŠŸ¨¥´¥´ÊŠ„µ¦Á„·— somatic embryos Á¤ºÉ°Áž¦¸¥Áš¸¥„´ »—‡ª‡» ¨³¡ªnµ ­¼˜¦°µ®µ¦ 1/2 MS š¸Éž¦³„°—oª¥­µ¦‡ª‡»¤ „µ¦Á˜·Ã˜ÅŽÃ˜Å‡œ·œ œ·— TDZ ‡ªµ¤Á o¤ oœ 1 ¤·¨¨·„¦´¤˜n°¨·˜¦ ­µ¤µ¦™´„œÎµÄ®oÁ„·— somatic embryos ¤µ„š¸É­»— š¸É¦·Áª– ž¨µ¥Ä adaxial side ¨³µ—Ÿ¨ ‡·—Áž}œ 75, 40 ¨³ 15 Áž°¦rÁŽÈœ˜r ˜µ¤¨Îµ—´ ¨³¤¸‹Îµœªœ somatic embryos ÁŒ¨¸É¥ 10.7 somatic embryos ˜n°·Êœ­nªœÄš¸Éš—¨°Š Park et al. (2002a) «¹„¬µ„µ¦ ¥µ¥¡´œ›»r„¨oª¥Å¤ o Phalaenopsis 4 ­µ¥¡´œ›»r ‡º° Tinny Sunshine ‘Annie’, ‘Taisuco Hatarot’, Teipei Gold ‘Golden Star’ ¨³ Tinny Galaxy ‘Annie’ —oª¥ ª·›¸„µ¦Á¡µ³Á¨¸Ê¥ŠÁœºÊ°Á¥ºÉ°‹µ„·Êœ­nªœÄ ¡ªnµ ­¼˜¦°µ®µ¦ 1/2 MS š¸Éž¦³„°—ª¥­µ¦‡ª‡o »¤„µ¦ Á˜·Ã˜ BA ‡ªµ¤Á o¤ oœ 88.8 ŤÇ¦Ã¤¨µ¦r ¦nª¤„´ NAA ‡ªµ¤Á o¤ oœ 5.4 ŤÇ¦Ã¤¨µ¦r ­µ¤µ¦™ ´„œÎµÄ®oÁ„·— PLBs ¤µ„š¸É­»— 85 Áž°¦rÁŽÈœ˜r ¨³¤¸‹Îµœªœ PLBs ÁŒ¨¸É¥ 10-13 PLBs ˜n°·Êœ­nªœÄš¸É š—¨°Š ®¨´Š‹µ„Á¡µ³Á¨¸Ê¥ŠÁžœÁª¨µ} 12 ­´ž—µ®r ¨³¡ªnµ PLBs Á¡·É¤‹µœªœ°¥Î nµŠ¤µ„Á¤ºÉ°Á¡µ³Á¨¸Ê¥Š œ­¼˜¦°µ®µ¦—´—ž¨Š Hyponex (1 „¦´¤˜n°¨·˜¦ 6.5N-4.5P-19K + 20N-20P-20K + ÁžžÃ˜œ 2 „¦´¤˜n°¨·˜¦ + œÊε˜o¤¤´œ ¦´ÉŠ 3 Áž°¦rÁŽÈœ˜r + ŸŠ™nµœ„¤¤´ ´œ˜r 0.05 Áž°¦rÁŽÈœ˜r) ¤¸‹Îµœªœ PLBs Á„·— ¹ÊœÁŒ¨¸É¥ 13-18 PLBs ˜n° 1 PLB š¸Éš—¨°Š 20

Park et al. (2002b) «¹„¬µ„µ¦ ¥µ¥¡œ›´ »r„¨oª¥Å¤o Doritaenopsis hybrid —oª¥ª·›¸„µ¦ Á¡µ³Á¨¸Ê¥ŠÁœºÊ°Á¥ºÉ°‹µ„œ­·Ê nªœÄ —oª¥ª·›¸ thin-section culture Á¡ºÉ°´„œÎµÄ®oÁ„·— PLBs ×¥˜¦Š ¡ªnµ ­¼˜¦°µ®µ¦ 1/2 MS š¸Éž¦³„°—oª¥­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ TDZ ­nŠŸ¨˜n°„µ¦´„œÎµÄ®oÁ„·— PLBs ¤µ„„ªnµ­¼˜¦°µ®µ¦ 1/2 MS š¸Éž¦³„°—ª¥­µ¦‡ª‡o »¤„µ¦Á˜·Ã˜ BA ¨³ zeatin Á¤ºÉ°Á¡µ³Á¨¸Ê¥Š ·Êœ­nªœÄ®œµ 1 ¤·¨¨·Á¤˜¦ œ­¼˜¦°µ®µ¦ 1/2 MS š¸Éž¦³„°—ª¥­µ¦‡ª‡o »¤„µ¦Á˜·Ã˜ TDZ ‡ªµ¤Á o¤ oœ 9.0 ŤÇ¦Ã¤¨µ¦r ¤¸‹Îµœªœ PLBs Á„·— ¹Êœ 72.3 Áž°¦rÁŽÈœ˜r ¨³¤¸‹Îµœªœ PLBs ÁŒ¨¸É¥ 18 PLBs ˜n°·Êœ­nªœÄš¸Éš—¨°Š Ž¹ÉŠ PLBs Á„·—Ä„¨o„´¦Áª–µ—Ÿ¨Â¨³­· nªœ epidermal cell Chen and Chang (2004) «¹„¬µŸ¨ °Š­µ¦‡ª‡»¤„µ¦Á˜Ã˜· 2,3,5-triiodobenzoic acid (TIBA) ˜n°„µ¦´„œÎµÄ®Á„o ·— somatic embryos ×¥˜¦Š ‹µ„·Êœ­ªœÄ °Š„¨n oª¥Å¤ o Oncidium Gower Ramsey —oª¥ª·›¸„µ¦Á¡µ³Á¨¸Ê¥ŠÁœºÊ°Á¥ºÉ° ¡ªnµ ­¼˜¦°µ®µ¦ 1/2 MS š¸Éž¦³„°—oª¥­µ¦‡ª‡»¤ „µ¦Á˜·Ã˜ TIBA ‡ªµ¤Á o¤ oœ 0.5 ŤÇ¦Ã¤¨µ¦r ­µ¤µ¦™´„œÎµÄ®oÁ„·— somatic embryos š¸É¦·Áª– ž¨µ¥Ä adaxial side, abaxial side ¨³¦°¥˜´— ‡·—Áž}œ 55, 47.5, 27.5 ¨³ 20 Áž°¦rÁŽÈœ˜r ˜µ¤¨Îµ—´

¨³¤¸‹Îµœªœ somatic embryos ÁŒ¨¸É¥ 134.2 embryos ˜n°‹µœÁ¡µ³Á¨¸Ê¥Š ĜÁª¨µ 8 ­´ž—µ®r ¨³¡ªnµ ­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ IAA, 2,4-D, 2-(3,4-dihydroxyphenyl)-3,5,7- trihydroxy-4H-chromen-4-one (quercetin) ¨³ 2-(p-chlorophenoxy)-2-methylpropionic acid (PICB) ­nŠŸ¨¥´¥´ÊŠ„µ¦Á„·— somatic embryos

Chen and Chang (2006) «¹„¬µ„µ¦ ¥µ¥¡´œ›»r„¨oª¥Å¤ o Phalaenopsis amabilis —oª¥

ª·›¸„µ¦Á¡µ³Á¨¸Ê¥ŠÁœºÊ°Á¥ºÉ°‹µ„·Êœ­nªœÄ Á¡°ºÉ ´„œÎµÄ®oÁ„·— somatic embryos ×¥˜¦Š‹µ„ epidermal cell ¡ªnµ ­¼˜¦°µ®µ¦ 1/2 MS š¸Éž¦³„°—oª¥­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ TDZ ‡ªµ¤Á o¤ oœ 3.0 ŤÇ¦ 䨵¦r ­µ¤µ¦™´„œÎµÄ®oÁ„·— somatic embryos ¤µ„š¸É­»— ÁŒ¨¸É¥ 19.4 embryos ˜n°·Êœ­nªœÄš¸É š—¨°Š Kuo et al. (2005) «¹„¬µ„µ¦ ¥µ¥¡œ›´ »r„¨oª¥Å¤o Phalaenopsis Little Steve —oª¥ª·›¸„µ¦ Á¡µ³Á¨¸Ê¥ŠÁœºÊ°Á¥ºÉ°‹µ„œ­·Ê nªœÄ Á¡ºÉ°´„œÎµÄ®oÁ„·— somatic embryos ×¥˜¦Š ¡ªnµ ­¼˜¦°µ®µ¦ 1/2 MS š¸Éž¦³„°—oª¥­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ BA ¨³ TDZ ­µ¤µ¦™´„œÎµÄ®oÁ„·— somatic embryos š¸É ¦·Áª–ŸªÄÂ¨³µ—Ÿ¨· ®¨´Š‹µ„Á¡µ³Á¨¸Ê¥ŠÄœš¸É¤º—ÁžœÁª¨µ} 20-30 ª´œ ¨³¡ªnµ ­¼˜¦°µ®µ¦š¸É ž¦³„°—ª¥­µ¦‡ª‡o »¤„µ¦Á˜·Ã˜ 2,4-D ‡ªµ¤Á o¤ oœ­¼Š ­nŠŸ¨¥´¥´ÊŠ„µ¦Á„·— somatic embryos Chung et al. (2007) «¹„¬µ„µ¦ ¥µ¥¡œ›´ »r„¨oª¥Å¤o Dendrobium cv. Chiengmai Pink —oª¥ª·›¸„µ¦Á¡µ³Á¨¸Ê¥ŠÁœºÊ°Á¥ºÉ°‹µ„·Êœ­nªœÄ œµ— 1 ÁŽœ˜·Á¤˜¦ Á¡ºÉ°´„œÎµÄ®oÁ„·— somatic embryos ×¥˜¦Š ¡ªnµ ­¼˜¦°µ®µ¦ 1/2 MS š¸Éž¦³„°—ª¥­µ¦‡ª‡o »¤„µ¦Á˜·Ã˜ TDZ ‡ªµ¤Á o¤ oœ 1 ¤·¨¨·„¦´¤˜n°¨·˜¦ Á®¤µ³­¤˜n°„µ¦´„œÎµÄ®oÁ„·— somatic embryos ¤µ„š¸É­»— ®¨´Š‹µ„Á¡µ³Á¨¸Ê¥ŠÁž}œ 21

Áª¨µ 60 ª´œ somatic embryos ­nªœÄ®nÁ„·— ¹Êœš¸É¦·Áª–ŸªÄÄ„¨· o„´¦·Áª–¦°¥˜´—¨³¦·Áª– ž¨µ¥ÄÁ¨È„œo°¥ Gow et al. (2008) «¹„¬µŸ¨ °Š­µ¦‡ª‡»¤„µ¦Á˜·Ã˜˜n°„µ¦´„œÎµÄ®oÁ„·— embryo ×¥˜¦Š‹µ„œ­·Ê nªœÄ„¨oª¥Å¤o Phalaenopsis 2 ­µ¥¡´œ›»r ‡º° Phalaenopsis amabilis ¨³ Phalaenopsis Nebula ¡ªnµ ¡´œ›»„¦¦¤¤¸Ÿ¨˜n°„µ¦´„œÎµÄ®oÁ„·— embryo ­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ BA ‡ªµ¤Á o¤ oœ 13.3 ŤÇ¦Ã¤¨µ¦r ¨³ 6-(Ȗ,Ȗ-Dimethylallylamino) purine (2iP) ‡ªµ¤Á o¤ oœ 4.92 ŤÇ¦Ã¤¨µ¦r ¤¸‡ªµ¤Á®¤µ³­¤˜n°„µ¦´„œÎµÄ®oÁ„·— embryo „´­µ¥¡´œ›»r Phalaenopsis amabilis ¨³ Phalaenopsis Nebula ˜µ¤¨Îµ—´

10. „µ¦Á¡µ³Á¨¸Ê¥ŠÁœºÊ°Á¥ºÉ°„¨oª¥Å¤o‹µ„ÁœºÊ°Á¥ºÉ°¦³®ªnµŠ o° °Š„oµœn°—°„Äœ­£µ¡ž¨°—ÁºÊ° ‹µ„¦µ¥Šµœª·‹´¥„µ¦Á¡µ³Á¨¸Ê¥ŠÁœºÊ°Á¥ºÉ°„¨oª¥Å¤o ¤¸¦µ¥Šµœ„µ¦Á¡µ³Á¨¸Ê¥ŠÁœºÊ°Á¥ºÉ°‹µ„ ÁœºÊ°Á¥ºÉ°¦³®ªnµŠ o°‡n°œ oµŠœo°¥ ¡°­¦»žÅ—o—´Šœ ¸Ê

Lin (1986) «¹„¬µ„µ¦ ¥µ¥¡´œ›»r„¨oª¥Å¤ o Phalaenopsis ¨³ Doritaenopsis ‹µ„„µ¦ Á¡µ³Á¨¸Ê¥ŠÁœºÊ°Á¥ºÉ°¦³®ªnµŠ o° °Š„oµœn°—°„ œ­¼˜¦°µ®µ¦ VW š¸Éž¦³„°—ª¥­µ¦‡ª‡o »¤„µ¦ Á˜·Ã˜ BA ‡ªµ¤Á o¤ oœ 1 ¨³ 5 ¤·¨¨·„¦´¤˜n°¨·˜¦ ¡ªnµ ¤¸„µ¦Á„·— PLBs ‹µ„ÁœºÊ°Á¥ºÉ°­nªœÁ°¡·Á—°¦r ¤·­ ¨³Ÿ·ª®œoµ °Š¦°¥˜´— ¨³¡´•œµÁž}œ˜oœš¸É­¤¦–¼ r®¨´Š‹µ„Á¡µ³Á¨¸Ê¥ŠÁžœÁª¨µ} 60 ª´œ ¨³

š¦µªnµ¦³¥³„µ¦¡´•œµ °Š„oµœn°—°„¤¸‡ªµ¤­Îµ‡´˜n°„µ¦´„œÎµÄ®oÁ„·— PLBs Ž¹ÉŠ¦³¥³š¸É

Á®¤µ³­¤‡º°„°œ„µ¦Á„n ·——°„¦„ ¨³ÁœºÊ°Á¥ºÉ°¦·Áª–Ä„¨ož¨µ¥¥°— °Š„oµœn°—°„¤¸‡ªµ¤ Á®¤µ³­¤š¸É­»—˜n°„µ¦´„œÎµ„µ¦Á„·— PLBs Chen et al. (2002) «¹„¬µ„µ¦ ¥µ¥¡œ›´ »r„¨oª¥Å¤o Epidendrum radicans ‹µ„„µ¦ Á¡µ³Á¨¸Ê¥ŠÁœºÊ°Á¥ºÉ°¦³®ªnµŠ o° °Š„oµœn°—°„ œ­¼˜¦°µ®µ¦ 1/2 MS š¸Éž¦³„°—oª¥­µ¦‡ª‡»¤ „µ¦Á˜·Ã˜ TDZ ‡ªµ¤Á o¤ oœ 0.45 ŤÇ¦Ã¤¨µ¦r ¡ªnµ ­µ¤µ¦™´„œÎµ„µ¦Á„·— PLBs ĜÁª¨µ 2 Á—º°œ ¨³­µ¤µ¦™Á¡·É¤‹Îµœªœ PLBs ŗo¤µ„š¸É­»— 23.6 PLBs Ĝ­¼˜¦°µ®µ¦š¸Éž¦³„°—oª¥œÊ嘵¨

Ž¼Ã‡¦­ ‡ªµ¤Á o¤ oœ 40 „¦´¤˜n°¨·˜¦ NH4NO3 ‡ªµ¤Á o¤ oœ 825 ¤·¨¨·„¦´¤˜n°¨·˜¦ ¨³ KNO3 ‡ªµ¤ Á o¤ oœ 950 ¤·¨¨·„¦´¤˜n°¨·˜¦ ¨³ PLBs ­µ¤µ¦™¡´•œµÁž}œ˜oœš­¤¸É ¼¦–rÁ¤ºÉ°Á¡µ³Á¨¸Ê¥Šœ­¼˜¦ °µ®µ¦š¸Éž¦³„°—oª¥­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ TDZ š¸É‡ªµ¤Á o¤ oœÁ—·¤ ĜÁªÁ¨µ 1 Á—º°œ

22

11. „µ¦«¹„¬µŸ¨ °ŠœÊ嘵¨Äœ„µ¦Á¡µ³Á¨¸Ê¥ŠÁœºÊ°Á¥ºÉ°„¨oª¥Å¤oĜ­£µ¡ž¨°—ÁºÊ° ‹µ„¦µ¥Šµœª·‹´¥„µ¦Á¡µ³Á¨¸Ê¥ŠÁœºÊ°Á¥ºÉ°„¨oª¥Å¤o¤¸Ÿ¼oª·‹´¥®¨µ¥šnµœÅ—oš—¨°Š«¹„¬µŸ¨ °Š œÊ嘵¨œ·—˜nµŠÇ ˜n°„µ¦Á‹¦·Á˜·Ã˜ °Š„¨oª¥Å¤o Á¡°Ä®ºÉ o„µ¦ ¥µ¥¡´œ›»r„¨oª¥Å¤o¤¸ž¦³­·š›·£µ¡ ¤µ„ ¹Êœ ¡°­¦»žÅ—o—´Šœ ¸Ê Islam and Ichihashi (1999) «¹„¬µœÊ嘵¨ 3 œ·— (Ž¼Ã‡¦­ ¤°¨Ã˜­ ¨³Ž°·š°¨) ˜n° „µ¦¡´•œµ °ŠÂ‡¨¨´­Â¨³„µ¦Á¡·É¤‹Îµœªœ °Š„¨oª¥Å¤ o Phalanopsis, Doritaenopsis ¨³ Neofinetia ¡ªnµ ‡¨¨´­ °Š„¨oª¥Å¤oš´ÊŠ 3 œ·— š¸ÉÁ¡µ³Á¨¸Ê¥Šœ°µ®µ¦š¸É¤¸œÊ嘵¨Ž¼Ã‡¦­¤¸„µ¦ ¡´•œµ¤µ„ ¹ÊœÁ¤ºÉ°Ä­nœÊ夳¡¦oµªÂ˜n¤¸­¸Á®¨°Šº Á„µ³„œÂ®¨ª¤Ç´ ®¨´Š‹µ„¥oµ¥ÅžÁ¡µ³Á¨¸Ê¥Šœ °µ®µ¦Ä®¤n 3 ‡¦´ÊŠ ‡¨¨´­š¸ÉÁ¡µ³Á¨¸Ê¥Šœ°µ®µ¦š¸É¤¸œÊ嘵¨¤°¨Ã˜­ ®¦º°Ž°·š°¨­nªœÄ®n ¡´•œµ Áž}œ PLBs ¨³¥´Š¡´•œµÅžÁž}œ˜oœš¸É­¤¦–¼ r ž„˜·Â¨³Â Ȋ¦Š ¤¸Â‡¨¨´­ °Š„¨oª¥Å¤o Phalanopsis ¨³ Neofinetia ­nªœœo°¥š¸É¡´•œµÁž}œÂ‡¨¨´­­¸Á ¸¥ªÂ¨³¡´•œµ˜n°Åž ˜nŤnÁ„—Áž· }œ PLBs ®¦º°¡º š¸É­¤¼¦–r

Tokuhara and Mii (2003) «¹„¬µœÊ嘵¨ 6 œ·— („¨¼Ã‡­ ¢¦»‡Ã˜­ ¤°¨Ã˜­ ¨„Ø­ Ž°·š°¨ ¨³ Ž¼Ã‡¦­ ‡ªµ¤Á o¤ oœ 58.4 ¤·¨¨·Ã¤¨µ¦r) ˜n°„µ¦´„œÎµÄ®oÁ„·— somatic embryos ‹µ„ „µ¦Á¡µ³Á¨¥ŠÁŽ¨¨¸Ê r ªœ¨°¥ °Š„¨oª¥Å¤ o Doriataenopsis —oª¥ª·›¸„µ¦Á¡µ³Á¨¸Ê¥ŠÁœ°Á¥ºÊ ºÉ° œ­¼˜¦ °µ®µ¦Â Ȋ New Dogashima medium (NDM) š¸Éž¦³„°—oª¥­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ NAA ‡ªµ¤

Á o¤ oœ 0.5 ŤÇ¦Ã¤¨µ¦r ¦nª¤„´ BA ‡ªµ¤Á o¤ oœ 4.4 ŤÇ¦Ã¤¨µ¦r ¡ªnµ ­¼˜¦°µ®µ¦š¸É

ž¦³„°—ª¥œo Ê嘵¨„¨¼Ã‡­ ­µ¤µ¦™´„œÎ µÄ®oÁ„·— PLBs ŗo—¸š¸É­»— œÊ嘵¨¤°¨Ã˜­ ¨³Ž°·š°¨ ­µ¤µ¦™´„œÎµÄ®oÁ„·— PLBs ˜nŤn¡„µ¦Á¡·É¤ž¦·¤µ– °ŠÂ‡¨¨´­ œÊ嘵¨Ž¼Ã‡¦­­µ¤µ¦™´„œÎµÄ®o Á„·—‡¨¨´­Å—Á®¤o º°œ„´œÊ嘵¨„¨¼Ã‡­ ˜nŤn­µ¤µ¦™´„œÎµÄ®oÁ„·— PLBs œÊ嘵¨¢¦»‡Ã˜­­µ¤µ¦™ Á¡·É¤ž¦·¤µ–‡¨¨´­Å—ož¦³¤µ–‡¦¹ÉŠ®œ¹ÉŠ °ŠœÊ嘵¨„¨¼Ã‡­ œÊ嘵¨Â¨„حŤn­µ¤µ¦™´„œÎµÄ®o Á„·—š´ÊŠ PLBs ¨³Â‡¨¨´­ Faria et al. (2004) «¹„¬µ‡ªµ¤Á o¤ oœ °ŠœÊ嘵¨Ž¼Ã‡¦­ (0, 5, 10, 20, 30 ¨³ 60 „¦´¤ ˜n°¨·˜¦) ˜n°„µ¦Á‹¦·Á˜·Ã˜Â¨³„µ¦Á„·—¦µ„„¨oª¥Å¤o Dendrobium nobile —oª¥ª·›¸„µ¦Á¡µ³Á¨¸Ê¥Š ÁœºÊ°Á¥ºÉ° œ­˜¦°µ®µ¦—¼ ´—ž¨Š MS ¡ªnµ ­¼˜¦°µ®µ¦——ž¨Š´ MS š¸É¤¸œÊ嘵¨Ž¼Ã‡¦­‡ªµ¤Á o¤ oœ 60 „¦´¤˜n°¨·˜¦ ­nŠŸ¨Ä®o˜oœ°n°œ¤¸‡ªµ¤­¼Š¤µ„š¸É­»—ÁŒ¨¸É¥ 4.21 ± 0.60 ÁŽœ˜·Á¤˜¦ ¨³¤¸„µ¦Á„—¥°—· šª¸‡¼–Äœ°´˜¦µ­nªœ 1 ˜n° 4 ˜nœÊ嘵¨Ž¼Ã‡¦­Å¤n¤¸°·š›·Ÿ¨˜n°„µ¦Á„·—¦µ„ °Š˜oœ°n°œ Rahman et al. (2005) «¹„¬µœÊ嘵¨ 3 œ·— (Ž¼Ã‡¦­ ¤°¨Ã˜­ ‡ªµ¤Á o¤ oœ 20 „¦´¤˜n° ¨·˜¦ ¨³Ž°·š°¨ ‡ªµ¤Á o¤ oœ 10 „¦´¤˜n°¨·˜¦) ˜n°„µ¦Á¡·É¤ž¦·¤µ–‡¨¨´­Â¨³„µ¦´„œÎµÄ®oÁ„·— ˜oœš¸É­¤¼¦– °Š„¨r oª¥Å¤ o Doritaenopsis —oª¥ª·›¸„µ¦Á¡µ³Á¨¸Ê¥ŠÁœºÊ°Á¥ºÉ°œ­¼˜¦°µ®µ¦ New 23

Phalaenopsis (NP) ¡ªnµ ‡¨¨´­š¸ÉÁ¡µ³Á¨¸Ê¥Šœ­¼˜¦°µ®µ¦ NP š¸É¤¸œÊ嘵¨Ž¼Ã‡¦­ ­µ¤µ¦™Á¡·É¤ ž¦·¤µ–‡¨¨´­Å—o¤µ„„ªnµ­˜¦°µ®µ¦¼ NP š¸É¤¸œÊ嘵¨¤°¨Ã˜­Â¨³Ž°¦r·š°¨ ÁŽ¨¨r¤¸¨´„¬–³Á„µ³ „´œ°¥nµŠ®¨ª¤Ç ¤¸­¸Á®¨º°ŠÄ­ ­Š­n°ŠŸnµœÅ—oœo°¥ Ž¹ÉŠÁ®¤º°œ¨´„¬–³Â‡¨¨´­Á¦·É¤˜oœÂ¨³Å¤nÁ„·— PLBs ˜n‡¨¨´­š¸ÉÁ¡µ³Á¨¸Ê¥Šœ­¼˜¦°µ®µ¦ NP š¸É¤¸œÊ嘵¨¤°¨Ã˜­ ¤¸­¸Á ¸¥ªÂ¨³¤¸„µ¦Á„·—Áž}œ PLBs ¤µ„ªnµœÊ嘵¨Ž°¦r·š°¨ œÊ嘵¨Ž°¦r·š°¨ ­nŠŸ¨Ä®o PLBs ¡´•œµÁž}œ˜oœš­¤¸É ¼¦–rŗo—¸„ªnµ œÊ嘵¨¤°¨Ã˜­Â¨³Ž¼Ã‡¦­ ˜n°¥nµŠÅ¦„˜µ¤È œÊ嘵¨¤°¨Ã˜­šÎµÄ®o˜oœ°n°œ¤¸„µ¦Á‹¦·Á˜·Ã˜Â¨³ ¡´•œµÅ—o—¸„ªµœn Ê嘵¨Ž°¦r·š°¨Â¨³Ž¼Ã‡¦­ Jheng et al. (2006) «¹„¬µœ·—¨³‡ªµ¤Á o¤ oœ °ŠœÊ嘵¨š¸É­nŠŸ¨˜n°„µ¦Á‹¦·Á˜·Ã˜ °Š„¨oª¥Å¤ o Oncidium Gower Ramsey ¡ªnµ ¡ºš¸ÉÁ¡µ³Á¨¸Ê¥Šœ­¼˜¦°µ®µ¦ 1/2 MS š¸É ž¦³„°—ª¥œo Ê嘵¨¤°¨Ã˜­ ®¦º°š¦¸±µÃ¨­ ­nŠŸ¨Ä®o¡º¤¸„µ¦Á‹¦·Á˜·Ã˜¤µ„„ªµ¡n ºš¸ÉÁ¡µ³Á¨¸Ê¥Š œ­¼˜¦°µ®µ¦ 1/2 MS š¸Éž¦³„°—ª¥œo Ê嘵¨Ž¼Ã‡¦­ °¥nµŠ¤¸œ´¥­Îµ‡´ PLBs ‹Îµœªœ 6,000 PLBs ×¥ž¦³¤µ–Å—o‹µ„„µ¦Á¡µ³Á¨¸Ê¥ŠÂ‡¨¨´­Á¦·É¤˜oœš¸É¤¸œÊ宜´„ 1 „¦´¤ Ĝ¦³¥³Áª¨µ 2 Á—º°œ ¨³

¤µ„„ªnµ 50 Áž°¦rÁŽÈœ˜r °Š PLBs ¡´•œµÁž}œ¡ºš¸É­¤¼¦–rĜ¦³¥³Áª¨µ 4 Á—º°œ ®¨´Š‹µ„¥oµ¥Åž Á¡µ³Á¨¸Ê¥Šœ­¼˜¦°µ®µ¦š¸É¤¸œÊ嘵¨š¦¸±µÃ¨­ ž¦·¤µ– 20 „¦´¤˜n°¨·˜¦ Novotna et al. (2007) «¹„¬µœ·— °ŠœÊ嘵¨Â¨³­µ¦‡ª‡»¤„µ¦Á˜·Ã˜š¸É­nŠŸ¨˜n°„µ¦ Á‹¦·Á˜·Ã˜ °Š„¨oª¥Å¤o Dactylorhiza species ¡ªnµ œÊ嘵¨„¨¼Ã‡­‡ªµ¤Á o¤ oœ 10 „¦´¤˜n°¨·˜¦

¦nª¤„´ œÊ嘵¨Ž¼Ã‡¦­‡ªµ¤Á o¤ oœ 10 „¦´¤˜n°¨·˜¦ ­nŠŸ¨„¦³˜»oœÄ®o˜oœ°n°œ¤¸„µ¦¡•œµš´ ¸É—¸ °´˜¦µ

„µ¦Á‹¦·Á˜·Ã˜Â¨³‡ªµ¤¥µª °Š¥°—¤ „µ¦¡¸ ´•œµ—¸ ¹ÊœÁ¤ºÉ°Á¡µ³Á¨¸Ê¥Š˜œ°o n°œÄœ­¼˜¦°µ®µ¦š¸É¤¸­µ¦ ‡ª‡»¤„µ¦Á˜Ã˜· BA ¦nª¤„´ IBA ¨³°´˜¦µ„µ¦Á‹¦·Á˜·Ã˜Â¨³‡ªµ¤¥µª °Š¦µ„¤¸„µ¦¡´•œµ—¸ ¹ÊœÁ¤ºÉ°Á¡µ³Á¨¸Ê¥Š˜oœ°n°œÄœ­¼˜¦°µ®µ¦š¸É¤¸­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ IBA ¦nª¤„´ NAA Hong et al. (2008a) «¹„¬µŸ¨ °ŠœÊ嘵¨ 5 œ·— (ÁŽ¨Ã¨ÅÃ°­ ¢¦»‡Ã˜­ „¨¼Ã‡­ ¤°¨Ã˜­ ¨³Ž¼Ã‡¦­ ‡ªµ¤Á o¤ oœ 10, 20, 30 ¨³ 60 „¦´¤˜n°¨·˜¦) ˜n°„µ¦´„œÎµÄ®oÁ„·— somatic embryos ‹µ„·Êœ­nªœÄ °Š„¨oª¥Å¤o Oncidium 2 ­µ¥¡´œ›»r (Oncidium cv. Gower Ramsey ¨³ Oncidium cv. Sweet Sugar) ¡ªnµ ­¼˜¦°µ®µ¦ 1/2 MS š¸Éž¦³„°—ª¥œo Ê嘵¨ÁŽ¨Ã¨ÅÃ°­ ­nŠŸ¨ ¥´¥´ÊŠ„µ¦Á„·— somatic embryos ¨³šÎµÄ®Á„o ·— browning „´„¨oª¥Å¤oš´ÊŠ 2 ­µ¥¡´œ› »r Ĝ„¨oª¥Å¤o Oncidium cv. Gower Ramsey ¡ªnµ ­¼˜¦°µ®µ¦ 1/2 MS š¸Éž¦³„°—oª¥œÊ嘵¨Ž¼Ã‡¦­ ‡ªµ¤ Á o¤ oœ 60 „¦´¤˜n°¨·˜¦ ¤¸‡ªµ¤Á®¤µ³­¤˜n°„µ¦´„œÎµÄ®oÁ„·— somatic embryos ¤µ„š¸É­»— ¨³Äœ „¨oª¥Å¤o Oncidium cv. Sweet Sugar ¡ªnµ ­¼˜¦°µ®µ¦ 1/2 MS š¸Éž¦³„°—oª¥œÊ嘵¨„¨¼Ã‡­ ‡ªµ¤ Á o¤ oœ 20 „¦´¤˜n°¨·˜¦ ¤¸‡ªµ¤Á®¤µ³­¤˜°„µ¦n ´„œÎµÄ®Á„o ·— somatic embryos ¤µ„š¸É­»—

šš¸É 3

°»ž„¦–r ­µ¦Á‡¤¸Â¨³ª·›¸„µ¦š—¨°Š

1. °»ž„¦–r¨³­µ¦Á‡¤¸ 1.1 ¡ºš—¨°Š „¨oª¥Å¤o Paphiopedilum ‘Delrosi’ Ĝ ª—ž¨°—ÁºÊ° °µ¥ » 14 Á—º°œ®¨´Š‹µ„Á¡µ³Á¤¨È— ‹µ„ KIT’s Orchids 56/3 Ž°¥„¦»ŠÁš¡œœšr 7 ™œœ„¦»ŠÁš¡²-œœš»¦¸ °ÎµÁ£°Á¤º°Š ‹´Š®ª´—œœš»¦¸ ž¦³Áš«Åš¥ 1.2 ­µ¦Á‡¤¸š¸ÉčoĜ„µ¦Á˜¦¸¥¤°µ®µ¦

1.2.1 ­µ¦Á‡¤¸š¸ÉÁž}œ°Š‡rž¦³„° °Š°µ®µ¦­´ŠÁ‡¦µ³®r - Hyponex (N:P:K = 6.5-6-19; Hyponex Co., Marysville, Ohio, USA) - MS (Murashige and Skoog, 1962)

- Phytagel (Sigma, P 8169) - Peptone (Criterion, C6531)

- ŸŠ™nµœ„´¤¤œ˜´ r (activated charcoal) 1.2.2 ®¨nŠ‡µ¦rÝűÁ—¦˜ - Ž¼Ã‡¦­ (sucrose) - „¨¼Ã‡­ (glucose) - ¤°¨Ã˜­ (maltose) 1.2.3 ­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ - 2,4-dichlorophenoxyacetic acid (2,4-D) - 6-benzyladenine (BA) - adenine sulfate (Ads) - kinetin (Kn) - thidiazuron (TDZ)

24

25

1.3 °»ž„¦–r 1.3.1 °»ž„¦–r¨³ª´­—»­Îµ®¦´ÄoÁ˜¦¸¥¤°µ®µ¦Á¡µ³Á¨¸Ê¥Š - ®¤o°œ¹ÉŠ‡ªµ¤—´œÅ° (autoclave) - Á‡¦ºÉ°Š´ÉŠÂ¨³Á°¸¥—¨³®¥µ - Á‡¦ºÉ°Šª´—‡ªµ¤Áž}œ„¦—-—nµŠ (pH meter) - Á˜µÅ¢¢jµ (hot plate) - Á˜µÅ¤Ã‡¦Áª¢ (microwave) - o°œ˜´„­µ¦Á‡¤¸ - Á‡¦ºÉ°ŠÂ„oª: ¸„Á„°¦r ª—Ä­n°µ®µ¦ šnŠÂ„oª „¦³°„˜ªŠ ¨³ždÁž˜˜r 1.3.2 °»ž„¦–r¨³ª´­—»­Îµ®¦´˜´—¨³™nµ¥ÁœºÊ°Á¥ºÉ° - ˜¼o™nµ¥ÁœºÊ°Á¥ºÉ°¡º (lamina air flow) - ˜³Á„¸¥ŠÂ°¨„°±°¨r

- ‹µœÂ„oª - Á‡¦ºÉ°Š¤º°Ÿnµ˜´—: —oµ¤¤¸— ĝ¤¸— ¨³žµ„‡¸ - ª—nÁ‡¦ºÉ°Š¤º°

£µ¡š¸É 2 ˜oœ„¨oª¥Å¤o Paphiopedilum ‘Delrosi’ Ĝ ª—ž¨°—ÁºÊ° °µ¥» 14 Á—º°œ®¨´Š‹µ„Á¡µ³Á¤¨È—

26

2. ª·›¸„µ¦š—¨°Š 2.1 «¹„¬µ­¼˜¦°µ®µ¦š¸ÉÁ®¤µ³­¤˜n°„µ¦´„œÎµ„µ¦Á„·—¥°—šª¸‡¼–‹µ„­ªœ¥°— °Š„¨n oª¥Å¤ o Paphiopedilum ‘Delrosi’ 2.1.1 «¹„¬µŸ¨ °Š­µ¦‡ª‡¤„µ¦Á˜» ·Ã˜ TDZ, BA, Kn, 2,4-D ¨³ Ads ˜n°„µ¦´„œÎµ„µ¦ Á„·—¥°—šª¸‡¼–‹µ„­nªœ¥°— °Š„¨oª¥Å¤ o Paphiopedilum ‘Delrosi’ œÎµ˜oœ„¨ª¥Å¤o o Paphiopedilum ‘Delrosi’ °µ¥» 14 Á—º°œ®¨´Š‹µ„Á¡µ³Á¤¨È— ¤µÁ¡µ³Á¨¸Ê¥Š œ­¼˜¦°µ®µ¦—´—ž¨Š Hyponex [ž»l¥ Hyponex (6.5N-4.5P-19K) ‡ªµ¤Á ¤ o oœ 3 „¦´¤˜n° ¨·˜¦ + ÁžžÃ˜œ (peptone) ‡ªµ¤Á o¤ oœ 2 „¦´¤˜n°¨·˜¦ + œÊε˜o¤¤´œ ¦´ÉŠ 100 „¦´¤˜n°¨·˜¦] Áž}œÁª¨µ 2 ­´ž—µ®r (¦¼žš¸É 3A) Á¡ºÉ°Ä®o˜oœ„¨ª¥Å¤o ž¦o ´˜´ª„´­¼˜¦°µ®µ¦ oµŠ˜oœ ‹µ„œœœ´Ê 夵˜´—˜nŠÄÄ®o Á®¨º°‹Îµœªœ 3 ĝ ¨³˜´—¦µ„š¸ÉÁ„·— ¹Êœ (¦¼žš¸É 3B) ¨³Á¡µ³Á¨¸Ê¥Šœ­¼˜¦°µ®µ¦—´—ž¨Š Hyponex š¸Éž¦³„°—ª¥­µ¦‡ª‡o »¤„µ¦Á˜·Ã˜ —´Š˜µ¦µŠš¸É 1 šÎµ„µ¦š—¨°Š 10 ŽÊε ŽÊ娳 1 ª— Á¡µ³Á¨¥Šœ°µ®µ¦¸Ê  Ȋš¸ÉÁ˜·¤ª»oœ (Gelrite) 2.4 „¦´¤

˜n°¨·˜¦ œÊ嘵¨Ž¼Ã‡¦­ ‡ªµ¤Á o¤ oœ 20 „¦´¤˜n°¨·˜¦ ¨³ŸŠ™nµœ„´¤¤œ˜´ r ž¦·¤µ– 1 „¦´¤˜n°¨·˜¦ Ĝ ª—Á¡µ³Á¨¸Ê¥Šž¦·¤µ˜¦ 240 ¤·¨¨·¨·˜¦ š¸É¤¸°µ®µ¦ž¦·¤µ˜¦ 50 ¤·¨¨·¨·˜¦ (œ¹ÉŠ‰nµÁºÊ°š¸É°»–®£¼¤· 121 °Š«µÁŽ¨ÁŽ¸¥­ Áž}œÁª¨µ 15 œµš¸) pH 5.40±0.01 ­£µ¡š¸Éŗo¦´Â­Š­ªnµŠ‹µ„®¨°—¢¨¼°°Á¦­ÁŽœ˜rš¸É ¤¸‡ªµ¤Á o¤Â­Š 30-35 ŤÇ¦Ã¤¨˜n°˜µ¦µŠÁ¤˜¦˜n°ª·œµš¸ Áž}œÁª¨µ 16 ´ÉªÃ¤Š˜n°ª´œ °»–®£¼¤· 25±2

°Š«µÁŽ¨ÁŽ¸¥­

´œš¹„Ÿ¨®¨´Š„µ¦Á¡µ³Á¨¥ŠÁž¸Ê }œ¦³¥³Áª¨µ 16 ­´ž—µ® r ×¥´œš¹„œÊ宜´„­— ‹Îµœªœ ĝ¨³¦µ„ °Š¥°—„¨ª¥Å¤o Á¦o ·É¤˜oœ ¨³Áž°¦rÁŽÈœ˜r„µ¦Á„·—¥°— ‹Îµœªœ¥°— œÊ宜´„­— ‹ÎµœªœÄ ¨³¦µ„ °Š¥°—„¨oª¥Å¤oš¸ÉÁ„·—Ä®¤n

27

˜µ¦µŠš¸É 1 ­µ¦‡ª‡»¤„µ¦Á˜Ã˜· TDZ, BA, Kn, 2,4-D ¨³ Ads ˜n°„µ¦´„œÎµ„µ¦Á„—¥°—šª· ¸‡¼– ‹µ„­nªœ¥°— °Š„¨oª¥Å¤ o Paphiopedilum ‘Delrosi’

­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ ­¼˜¦°µ®µ¦ TDZ (ôM) BA (ôM) Kn (ôM) 2,4 -D (ôM) Ads (ôM) 1 2 0.45 3 4.54 4 2.22 5 4.44 6 4.65 7 9.29

8 0.45 4.52 9 4.54 4.52 10 2.22 4.52 11 4.44 4.52

12 4.65 4.52

13 0.45 27.10 14 4.54 27.10 15 2.22 27.10 16 4.44 27.10

28

£µ¡š¸É 3 ¨´„¬–³ °Š„¨ª¥Å¤o o Paphiopedilum ‘Delrosi’ (A) ˜oœ„¨oª¥Å¤oš¸ÉÁ¡µ³Á¨¸Ê¥Šœ­¼˜¦°µ®µ¦—´—ž¨Š Hyponex Áž}œÁª¨µ 2 ­´ž—µ® r

(B) ˜oœ„¨oª¥Å¤oš¸É˜´—˜nŠÄÄ®Á®¨o º°‹Îµœªœ 3 ĝ ¨³˜´—¦µ„°°„ (­nªœ¥°—)

29

2.1.2 «¹„¬µŸ¨ °Š­µ¦‡ª‡¤„µ¦Á˜» ·Ã˜ TDZ, BA, Kn ¨³ 2,4-D ˜n°„µ¦´„œÎµ„µ¦Á„·— ¥°—šª¸‡¼–‹µ„­nªœ¥°— °Š„¨oª¥Å¤ o Paphiopedilum ‘Delrosi’ œÎµ˜oœ„¨ª¥Å¤o o Paphiopedilum ‘Delrosi’ °µ¥» 19 Á—º°œ®¨´Š‹µ„Á¡µ³Á¤¨È— ¤µÁ¡µ³Á¨¸Ê¥Š œ­¼˜¦°µ®µ¦—´—ž¨Š Hyponex [ž»l¥ Hyponex (6.5N-4.5P-19K) ‡ªµ¤Á ¤ o oœ 3 „¦´¤˜n° ¨·˜¦ + ÁžžÃ˜œ (peptone) ‡ªµ¤Á o¤ oœ 2 „¦´¤˜n°¨·˜¦ + œÊε˜o¤¤´œ ¦´ÉŠ 100 „¦´¤˜n°¨·˜¦] ÁžœÁª¨µ} 2 ­´ž—µ®r (¦¼žš¸É 3A) Á¡ºÉ°Ä®o˜oœ„¨ª¥Å¤o ž¦o ´˜´ª„´­¼˜¦°µ®µ¦ oµŠ˜oœ ‹µ„œœœ´Ê 夵˜´—˜nŠÄÄ®o Á®¨º°‹Îµœªœ 3 ĝ ¨³˜´—¦µ„š¸ÉÁ„·— ¹Êœ (¦¼žš¸É 3B) ÁnœÁ—¸¥ª„´„µ¦š—¨°Š 2.1.1 ¨³Á¡µ³Á¨¸Ê¥Šœ ­¼˜¦°µ®µ¦—´—ž¨Š Hyponex š¸Éž¦³„°—ª¥­µ¦‡ª‡o »¤„µ¦Á˜·Ã˜ —´Š˜µ¦µŠš¸É 2 šÎµ„µ¦š—¨°Š 10 ŽÊε ŽÊ娳 1 ª— Á¡µ³Á¨¥Šœ°µ®µ¦Â ¸Ê Ȋš¸ÉÁ˜·¤ª»oœ (Gelrite) 2.4 „¦´¤ ˜n°¨·˜¦ œÊ嘵¨Ž¼Ã‡¦­ ‡ªµ¤Á o¤ oœ 20 „¦´¤˜n°¨·˜¦ ¨³ŸŠ™nµœ„´¤¤œ˜´ r ž¦·¤µ– 1 „¦´¤˜n°¨·˜¦ Ĝ ª—Á¡µ³Á¨¸Ê¥Šž¦·¤µ˜¦ 240 ¤·¨¨·¨·˜¦ š¸É¤¸°µ®µ¦ž¦·¤µ˜¦ 50 ¤·¨¨·¨·˜¦ (œ¹ÉŠ‰nµÁºÊ°š¸É°»–®£¼¤· 121 °Š«µÁŽ¨ÁŽ¸¥­ Áž}œÁª¨µ 15 œµš¸) pH 5.40±0.01 ­£µ¡š¸Éŗo¦´Â­Š­ªnµŠ‹µ„®¨°—¢¨¼°°Á¦­ÁŽœ˜r

š¸É¤¸‡ªµ¤Á o¤Â­Š 30-35 ŤÇ¦Ã¤¨˜n°˜µ¦µŠÁ¤˜¦˜n°ªœµš· ¸ Áž}œÁª¨µ 16 ´ÉªÃ¤Š˜n°ª´œ °»–®£¼¤· 25±2 °Š«µÁŽ¨ÁŽ¸¥­ ´œš¹„Ÿ¨®¨´Š„µ¦Á¡µ³Á¨¥ŠÁž¸Ê }œ¦³¥³Áª¨µ 16 ­´ž—µ® r ×¥´œš¹„œÊ宜´„­— ‹Îµœªœ ĝ¨³¦µ„ °Š¥°—„¨ª¥Å¤o Á¦o ·É¤˜oœ ¨³Áž°¦rÁŽÈœ˜r„µ¦Á„·—¥°— ‹Îµœªœ¥°— œÊ宜´„­— ‹ÎµœªœÄ

¨³¦µ„ °Š¥°—„¨oª¥Å¤oš¸ÉÁ„·—Ä®¤n

30

˜µ¦µŠš¸É 2 ­µ¦‡ª‡»¤„µ¦Á˜Ã˜· TDZ, BA, Kn ¨³ 2,4-D ˜n°„µ¦´„œÎµ„µ¦Á„—¥°—šª· ¸‡¼–‹µ„ ­nªœ¥°— °Š„¨oª¥Å¤ o Paphiopedilum ‘Delrosi’

­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ ­¼˜¦°µ®µ¦ TDZ (ôM) BA (ôM) Kn (ôM) 2,4 -D (ôM) 1 2 0.45 3 2.25 4 4.44 5 8.88 6 2.32

7 4.65 8 0.45 4.52 9 0.45 13.56 10 2.25 4.52

11 2.25 13.56

12 4.44 13.56 13 8.88 13.56

31

2.2 «¹„¬µœ·— °ŠœÊ嘵¨š¸ÉÁ®¤µ³­¤˜n°„µ¦´„œÎµ„µ¦Á„·—¥°—šª¸‡¼–‹µ„­nªœ¥°— °Š„¨oª¥Å¤o Paphiopedilum ‘Delrosi’ œÎµ˜oœ„¨ª¥Å¤o o Paphiopedilum ‘Delrosi’ °µ¥ » 14 Á—°œ®¨º ´Š‹µ„Á¡µ³Á¤¨È— ¤µ Á¡µ³Á¨¸Ê¥Šœ­¼˜¦°µ®µ¦—´—ž¨Š Hyponex [ž»l¥ Hyponex (6.5N-4.5P-19K) ‡ªµ¤Á o¤ oœ 3 „¦´¤˜n°¨·˜¦ + ÁžžÃ˜œ (peptone) ‡ªµ¤Á o¤ oœ 2 „¦´¤˜n°¨·˜¦ + œÊε˜o¤¤´œ ¦´ÉŠ 100 „¦´¤˜n°¨·˜¦] Áž}œÁª¨µ 2 ­´ž—µ®r (¦¼žš¸É 3A) Á¡ºÉ°Ä®o˜oœ„¨oª¥Å¤ož¦˜´ ´ª„´­¼˜¦°µ®µ¦ oµŠ˜oœ ‹µ„œ´ÊœœÎµ¤µ˜´— ˜nŠÄÄ®oÁ®¨º°‹Îµœªœ 3 ĝ ¨³˜´—¦µ„š¸ÉÁ„·— ¹Êœ (¦¼žš¸É 3B) ¨³Á¡µ³Á¨¸Ê¥Šœ­¼˜¦°µ®µ¦—´—ž¨Š Hyponex š¸Éž¦³„°—ª¥œo Ê嘵¨œ·—˜nµŠÇ ¨³­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ —´Š˜µ¦µŠš¸É 3 šÎµ„µ¦š—¨°Š 10 ŽÊε ŽÊ娳 1 ª— Á¡µ³Á¨¥Šœ°µ®µ¦Â ¸Ê Ȋš¸ÉÁ˜·¤ª»oœ (Gelrite) 2.4 „¦´¤ ˜n°¨·˜¦ œÊ嘵¨œ·—˜nµŠÇ ‡ªµ¤Á o¤ oœ 20 „¦´¤˜n°¨·˜¦ ¨³ŸŠ™nµœ„´¤¤´œ˜r ž¦·¤µ– 1 „¦´¤˜n°¨·˜¦ Ĝ ª—Á¡µ³Á¨¥Šž¦¸Ê ·¤µ˜¦ 240 ¤·¨¨·¨·˜¦ š¸É¤¸°µ®µ¦ž¦·¤µ˜¦ 50 ¤·¨¨·¨·˜¦ (œ¹ÉŠ‰nµÁºÊ°š¸É°»–®£¼¤· 121 °Š«µÁŽ¨ÁŽ¸¥­ Áž}œÁª¨µ 15 œµš¸) pH 5.40±0.01 ­£µ¡š¸Éŗo¦´Â­Š­ªnµŠ‹µ„®¨°—¢¨¼°°Á¦­ÁŽœ˜r

š¸É¤¸‡ªµ¤Á o¤Â­Š 30-35 ŤÇ¦Ã¤¨˜n°˜µ¦µŠÁ¤˜¦˜n°ªœµš· ¸ Áž}œÁª¨µ 16 ´ÉªÃ¤Š˜n°ª´œ °»–®£¼¤· 25±2 °Š«µÁŽ¨ÁŽ¸¥­ ´œš¹„Ÿ¨®¨´Š„µ¦Á¡µ³Á¨¥ŠÁž¸Ê }œ¦³¥³Áª¨µ 16 ­´ž—µ® r ×¥´œš¹„œÊ宜´„­— ‹Îµœªœ ĝ¨³¦µ„ °Š¥°—„¨ª¥Å¤o Á¦o ·É¤˜oœ ¨³Áž°¦rÁŽÈœ˜r„µ¦Á„·—¥°— ‹Îµœªœ¥°— œÊ宜´„­— ‹ÎµœªœÄ

¨³¦µ„ °Š¥°—„¨oª¥Å¤oš¸ÉÁ„·—Ä®¤n

32

˜µ¦µŠš¸É 3 œ·— °ŠœÊ嘵¨Â¨³­µ¦‡ª‡»¤„µ¦Á˜·Ã˜š¸ÉčoĜ„µ¦´„œÎµ„µ¦Á„·—¥°—šª¸‡¼–‹µ„­nªœ ¥°— °Š„¨oª¥Å¤o Paphiopedilum ‘Delrosi’

­¼˜¦ œÊ嘵¨ ­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ °µ®µ¦ 20 (g/l) TDZ (ôM) Kn (ôM) 2,4-D (ôM) 1 sucrose 0.45 4.52 2 sucrose 4.65 4.52 3 glucose 0.45 4.52 4 glucose 4.65 4.52 5 maltose 0.45 4.52 6 maltose 4.65 4.52

33

2.3 «¹„¬µ­¼˜¦°µ®µ¦š¸ÉÁ®¤µ³­¤˜n°„µ¦´„œÎµ„µ¦Á„·—¥°—šª¸‡¼–‹µ„˜œ„¨o oª¥Å¤o Paphiopedilum ‘Delrosi’ š¸É˜´—­nªœ¥°—°°„ œÎµ˜oœ„¨ª¥Å¤o o Paphiopedilum ‘Delrosi’ Á¡µ³Á¨¸Ê¥ŠÄœš¸É¤º—Áž}œÁª¨µ 16 ­´ž—µ®r Á¡ºÉ°Ä®o o° ÁœºÊ°Á¥ºÉ°¦³®ªnµŠ o° ¨³Äš¸É¦·Áª–¥°—¤¸‡ªµ¤¥µª¡°š¸É‹³˜—°°„Å—´ o ‹µ„œ´ÊœœÎµ˜oœš¸É˜´—­nªœ ¥°—°°„ (¦¼žš¸É 5) Á¡µ³Á¨¸Ê¥Šœ°µ®µ¦­ŠÁ‡¦µ³®´ r—´—ž¨Š­¼˜¦ Hyponex [ž»l¥ Hyponex (6.5N- 4.5P-19K) ‡ªµ¤Á o¤ oœ 3 „¦´¤˜n°¨·˜¦ + ÁžžÃ˜œ (peptone) ‡ªµ¤Á o¤ oœ 2 „¦´¤˜n°¨·˜¦ + œÊε˜o¤ ¤´œ ¦´ÉŠ 100 „¦´¤˜n°¨·˜¦] š¸Éž¦³„°—ª¥œo Ê嘵¨œ·—˜nµŠÇ ¨³­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ —´Š˜µ¦µŠš¸É 4 šÎµ„µ¦š—¨°Š 10 ŽÊε ŽÊ娳 1 ª— Á¡µ³Á¨¥Šœ°µ®µ¦¸Ê  Ȋš¸ÉÁ˜·¤ª»oœ (Gelrite) 2.4 „¦´¤ ˜n°¨·˜¦ œÊ嘵¨œ·—˜nµŠÇ ‡ªµ¤Á o¤ oœ 20 „¦´¤˜n°¨·˜¦ ¨³ŸŠ™nµœ„´¤¤´œ˜r ž¦·¤µ– 1 „¦´¤˜n°¨·˜¦ Ĝ ª—Á¡µ³Á¨¥Šž¦¸Ê ·¤µ˜¦ 240 ¤·¨¨·¨·˜¦ š¸É¤¸°µ®µ¦ž¦·¤µ˜¦ 50 ¤·¨¨·¨·˜¦ (œ¹ÉŠ‰nµÁºÊ°š¸É°»–®£¼¤· 121 °Š«µÁŽ¨ÁŽ¸¥­ Áž}œÁª¨µ 15 œµš¸) pH 5.40±0.01 ­£µ¡š¸Éŗo¦´Â­Š­ªnµŠ‹µ„®¨°—¢¨¼°°Á¦­ÁŽœ˜r š¸É¤¸‡ªµ¤Á o¤Â­Š 30-35 ŤÇ¦Ã¤¨˜n°˜µ¦µŠÁ¤˜¦˜n°ªœµš· ¸ Áž}œÁª¨µ 16 ´ÉªÃ¤Š˜n°ª´œ °»–®£¼¤·

25±2 °Š«µÁŽ¨ÁŽ¸¥­ ´œš¹„Ÿ¨®¨´Š„µ¦Á¡µ³Á¨¥ŠÁž¸Ê }œ¦³¥³Áª¨µ 16 ­´ž—µ®r ×¥´œš„„µ¦¦°—¹ ¸ª·˜ °Š˜oœ „¨oª¥Å¤oÁ—·¤ ¨³Áž°¦rÁŽÈœ˜r„µ¦Á„—¥°—· ‹Îµœªœ¥°— œÊ宜„­—´ ‹ÎµœªœÄÂ¨³¦µ„ °Š¥°— „¨oª¥Å¤oš¸ÉÁ„·—Ä®¤n

34

˜µ¦µŠš¸É 4 œ·— °ŠœÊ嘵¨Â¨³­µ¦‡ª‡»¤„µ¦Á˜·Ã˜š¸ÉčoĜ„µ¦´„œÎµ„µ¦Á„·—¥°—šª¸‡¼–‹µ„˜oœ „¨oª¥Å¤ o Paphiopedilum ‘Delrosi’ š¸É˜´—­nªœ¥°—°°„

œÊ嘵¨ ­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ ­¼˜¦°µ®µ¦ (20 g/l) TDZ (ôM) Kn (ôM) 2,4-D (ôM) 1 sucrose 2 sucrose 0.45 3 sucrose 0.45 4.52 4 sucrose 4.65 4.52 5 glucose 6 glucose 0.45 7 glucose 0.45 4.52

8 glucose 4.65 4.52 9 maltose 10 maltose 0.45 11 maltose 0.45 4.52

12 maltose 4.65 4.52

35

£µ¡š¸É 4 ¨´„¬–³˜œ„¨o oª¥Å¤o Paphiopedilum ‘Delrosi’ ®¨´Š‹µ„Á¡µ³Á¨¸Ê¥ŠÄœš¸É¤º— (A) ˜oœ„¨oª¥Å¤oš¸ÉÁ¡µ³Á¨¸Ê¥ŠÄœš¸É¤º— Áž}œÁª¨µ 16 ­´ž—µ® r

(B) ·Êœ­nªœÁœºÊ°Á¥ºÉ°˜µŠÇn š¸ÉœÎµ¤µÄÄœ„µ¦š—¨°Šo

£µ¡š¸É 5 ¨´„¬–³„µ¦Á¡µ³Á¨¸Ê¥Š˜oœ„¨ª¥Å¤o o Paphiopedilum ‘Delrosi’ š¸É˜´—­nªœ¥°—°°„

36

2.4 «¹„¬µ­¼˜¦°µ®µ¦š¸ÉÁ®¤µ³­¤˜n°„µ¦´„œÎµ„µ¦Á„·—¥°—‹µ„­nªœ o° °Š˜œ„¨o oª¥Å¤o Paphiopedilum ‘Delrosi’ œÎµ­nªœ o° °Š˜oœ„¨oª¥Å¤ o Paphiopedilum ‘Delrosi’ ‹µ„„µ¦š—¨°Šš¸É 2.3 Ş Á¡µ³Á¨¸Ê¥Šœ°µ®µ¦­´ŠÁ‡¦µ³®r—´—ž¨Š­¼˜¦ Hyponex [ž»l¥ Hyponex (6.5N-4.5P-19K) ‡ªµ¤ Á o¤ oœ 3 „¦´¤˜n°¨·˜¦ + ÁžžÃ˜œ (peptone) ‡ªµ¤Á o¤ oœ 2 „¦´¤˜n°¨·˜¦ + œÊε˜o¤¤´œ ¦´ÉŠ 100 „¦´¤ ˜n°¨·˜¦] (¦¼žš¸É 6) š¸Éž¦³„°—oª¥œÊ嘵¨œ—˜· nµŠÇ ¨³­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ —´Š˜µ¦µŠš¸É 5 šÎµ„µ¦š—¨°Š 10 ŽÊε ŽÊ娳 1 ª— Á¡µ³Á¨¥Šœ°µ®µ¦Â ¸Ê Ȋš¸ÉÁ˜·¤ª»oœ (Gelrite) 2.4 „¦´¤ ˜n°¨·˜¦ œÊ嘵¨œ·—˜nµŠÇ ‡ªµ¤Á o¤ oœ 20 „¦´¤˜n°¨·˜¦ ¨³ŸŠ™nµœ„´¤¤´œ˜r ž¦·¤µ– 1 „¦´¤˜n°¨·˜¦ Ĝ ª—Á¡µ³Á¨¥Šž¦¸Ê ·¤µ˜¦ 120 ¤·¨¨·¨·˜¦ š¸É¤¸°µ®µ¦ž¦·¤µ˜¦ 25 ¤·¨¨·¨·˜¦ (œ¹ÉŠ‰nµÁºÊ°š¸É°»–®£¼¤· 121 °Š«µÁŽ¨ÁŽ¸¥­ Áž}œÁª¨µ 15 œµš¸) pH 5.40±0.01 ­£µ¡š¸Éŗo¦´Â­Š­ªnµŠ‹µ„®¨°—¢¨¼°°Á¦­ÁŽœ˜r š¸É¤¸‡ªµ¤Á o¤Â­Š 30-35 ŤÇ¦Ã¤¨˜n°˜µ¦µŠÁ¤˜¦˜n°ªœµš· ¸ Áž}œÁª¨µ 16 ´ÉªÃ¤Š˜n°ª´œ °»–®£¼¤· 25±2 °Š«µÁŽ¨ÁŽ¸¥­

´œš¹„Ÿ¨®¨´Š„µ¦Á¡µ³Á¨¥ŠÁž¸Ê }œ¦³¥³Áª¨µ 16 ­´ž—µ® r ×¥´œš¹„Áž°¦rÁŽÈœ˜r„µ¦Á„·— ¥°— ‹Îµœªœ¥°— œÊ宜´„­— ‹ÎµœªœÄÂ¨³¦µ„ °Š¥°—„¨oª¥Å¤oš¸ÉÁ„—Ä®¤· n

37

˜µ¦µŠš¸É 5 œ·— °ŠœÊ嘵¨Â¨³­µ¦‡ª‡»¤„µ¦Á˜·Ã˜š¸ÉčoĜ„µ¦´„œÎµ„µ¦Á„·—¥°—‹µ„­nªœ o° °Š ˜oœ„¨oª¥Å¤ o Paphiopedilum ‘Delrosi’

œÊ嘵¨ ­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ ­¼˜¦°µ®µ¦ (20 g/l) TDZ (ôM) Kn (ôM) 2,4-D (ôM) 1 sucrose 2 sucrose 0.45 3 sucrose 0.45 4.52 4 sucrose 4.65 4.52 5 glucose 6 glucose 0.45

7 glucose 0.45 4.52 8 glucose 4.65 4.52 9 maltose 10 maltose 0.45

11 maltose 0.45 4.52

12 maltose 4.65 4.52

38

£µ¡š¸É 6 ¨´„¬–³„µ¦Á¡µ³Á¨¸Ê¥Š­nªœ o° °Š˜oœ„¨oª¥Å¤o Paphiopedilum ‘Delrosi’

39

2.5 «¹„¬µ­¼˜¦°µ®µ¦š¸ÉÁ®¤µ³­¤˜n°„µ¦´„œÎµ„µ¦Á„·—‡¨¨´­ ®¦º° somatic embryos ‹µ„·ÊœÁœºÊ° Á¥ºÉ°¦³®ªnµŠ o° °Š„¨oª¥Å¤ o Paphiopedilum ‘Delrosi’ œÎµ·ÊœÁœºÊ°Á¥ºÉ°¦³®ªnµŠ o° °Š˜oœ„¨oª¥Å¤o Paphiopedilum ‘Delrosi’ ‹µ„„µ¦š—¨°Šš¸É 2.3 ˜´—Ä®o¤¸‡ªµ¤¥µª 0.5 ÁŽœ˜·Á¤˜¦ ¨³œÎµÅžÁ¡µ³Á¨¥Šœ­¸Ê ¼˜¦°µ®µ¦ 1/2 MS (1/2 strength macro-and full-strength micro elements) š¸Éž¦³„°—ª¥œo µ˜µ¨œÊÎ ·—˜nµŠÇ ¨³­µ¦‡ª‡»¤„µ¦ Á˜·Ã˜ —´Š˜µ¦µŠš¸É 6 šÎµ„µ¦š—¨°Š 10 ŽÊε Á¡µ³Á¨¸Ê¥Šœ°µ®µ¦Â Ȋš¸ÉÁ˜·¤ª»oœ (Gelrite) 2.4 „¦´¤˜n°¨·˜¦ œÊ嘵¨œ·—˜nµŠÇ ‡ªµ¤Á o¤ oœ 20 „¦´¤˜n°¨·˜¦ Ĝ ª—Á¡µ³Á¨¸Ê¥Šž¦·¤µ˜¦ 120 ¤·¨¨·¨·˜¦ š¸É¤¸°µ®µ¦ ž¦·¤µ˜¦ 25 ¤·¨¨·¨·˜¦ (œ¹ÉŠ‰nµÁºÊ°š¸É°»–®£¼¤· 121 °Š«µÁŽ¨ÁŽ¸¥­ Áž}œÁª¨µ 15 œµš)¸ pH 5.40±0.01 ­£µ¡š¸Éŗo¦´Â­Š­ªnµŠ‹µ„®¨°—¢¨¼°°Á¦­ÁŽœ˜rš¸É¤¸‡ªµ¤Á o¤Â­Š 30-35 ŤÇ¦Ã¤¨˜n°˜µ¦µŠÁ¤˜¦ ˜n°ª·œµš¸ ÁžœÁª¨µ} 16 ´ÉªÃ¤Š˜n°ª´œ °–®£» ¼¤· 25±2 °Š«µÁŽ¨ÁŽ¸¥­ ´œš¹„Ÿ¨®¨´Š„µ¦Á¡µ³Á¨¥ŠÁž¸Ê }œ¦³¥³Áª¨µ 16 ­´ž—µ® r ×¥´œš¹„ ­¸ ¨´„¬–³ ¨³

ž¦·¤µ–‡¨¨´­ ®¦º° ­¸ ¨´„¬–³ ¨³‹Îµœªœ somatic embryos š¸ÉÁ„·— ¹Êœ

40

˜µ¦µŠš¸É 6 œ·— °ŠœÊ嘵¨Â¨³­µ¦‡ª‡»¤„µ¦Á˜·Ã˜š¸ÉčoĜ„µ¦´„œÎµ„µ¦Á„·—‡¨¨´­ ®¦º° somatic embryos ‹µ„œÁœ·Ê °Á¥ºÊ ºÉ°¦³®ªnµŠ ° °Š„¨o oª¥Å¤ o Paphiopedilum ‘Delrosi’

œÊ嘵¨ ­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ ­¼˜¦°µ®µ¦ (20 g/l) TDZ (ôM) 2,4-D (ôM) 1 sucrose 2 sucrose 0.45 3 sucrose 4.54 4 sucrose 0.45 4.52 5 sucrose 4.54 4.52 6 sucrose 4.54 13.56 7 glucose

8 glucose 0.45 9 glucose 4.54 10 glucose 0.45 4.52 11 glucose 4.54 4.52

12 glucose 4.54 13.56

13 maltose 14 maltose 0.45 15 maltose 4.54 16 maltose 0.45 4.52 17 maltose 4.54 4.52 18 maltose 4.54 13.56

41

£µ¡š¸É 7 ¨´„¬–³„µ¦Á¡µ³Á¨¸Ê¥Š·ÊœÁœºÊ°Á¥ºÉ°¦³®ªnµŠ o° °Š„¨oª¥Å¤o Paphiopedilum ‘Delrosi’

42

2.6 «¹„¬µ­¼˜¦°µ®µ¦š¸ÉÁ®¤µ³­¤˜n°„µ¦´„œÎµ„µ¦Á„·—‡¨¨´­ ®¦º° somatic embryos ‹µ„·Êœ­nªœ ĝ °Š„¨oª¥Å¤o Paphiopedilum ‘Delrosi’ œÎµÄÄœ­nªœ¥°— °Š˜oœ°°œ„¨n oª¥Å¤o Paphiopedilum ‘Delrosi’ ‹µ„„µ¦š—¨°Šš¸É 2.3 ˜´—ĝ°n°œ­nªœ‡¦¹ÉŠÄ¨nµŠÄ®o¤¸ œµ— 0.3 x 0.4 ˜µ¦µŠÁŽœ˜·Á¤˜¦ Á¡µ³Á¨¸Ê¥Šœ­¼˜¦°µ®µ¦ 1/2 MS (1/2 strength macro-and full-strength micro elements) š¸Éž¦³„°—ª¥­µ¦‡ª‡o »¤„µ¦Á˜·Ã˜ ¨³ œÊ嘵¨œ·—˜nµŠÇ —´Š˜µ¦µŠš¸É 7 šÎµ„µ¦š—¨°Š 10 ŽÊε Á¡µ³Á¨¸Ê¥Šœ°µ®µ¦Â Ȋš¸ÉÁ˜·¤ª»oœ (Gelrite) 2.4 „¦´¤˜n°¨·˜¦ œÊ嘵¨œ·—˜nµŠÇ ‡ªµ¤Á o¤ oœ 20 „¦´¤˜n°¨·˜¦ Ĝ ª—Á¡µ³Á¨¸Ê¥Šž¦·¤µ˜¦ 120 ¤·¨¨·¨·˜¦ š¸É¤¸°µ®µ¦ ž¦·¤µ˜¦ 25 ¤·¨¨·¨·˜¦ (œ¹ÉŠ‰nµÁºÊ°š¸É°»–®£¼¤· 121 °Š«µÁŽ¨ÁŽ¸¥­ Áž}œÁª¨µ 15 œµš)¸ pH 5.40±0.01 ­£µ¡š¸Éŗo¦´Â­Š­ªnµŠ‹µ„®¨°—¢¨¼°°Á¦­ÁŽœ˜rš¸É¤¸‡ªµ¤Á o¤Â­Š 30-35 ŤÇ¦Ã¤¨˜n°˜µ¦µŠÁ¤˜¦ ˜n°ª·œµš¸ ÁžœÁª¨µ} 16 ´ÉªÃ¤Š˜n°ª´œ °»–®£¼¤· 25±2 °Š«µÁŽ¨ÁŽ¸¥­ ´œš¹„Ÿ¨®¨´Š„µ¦Á¡µ³Á¨¥ŠÁž¸Ê }œ¦³¥³Áª¨µ 16 ­´ž—µ® r ×¥´œš¹„ ­¸ ¨´„¬–³ ¨³

ž¦·¤µ–‡¨¨´­ ®¦º° ­¸ ¨´„¬–³ ¨³‹Îµœªœ somatic embryos š¸ÉÁ„·— ¹Êœ

43

˜µ¦µŠš¸É 7 œ·— °ŠœÊ嘵¨Â¨³­µ¦‡ª‡»¤„µ¦Á˜·Ã˜š¸ÉčoĜ„µ¦´„œÎµ„µ¦Á„·—‡¨¨´­ ®¦º° somatic embryos ‹µ„œ­·Ê nªœÄ °Š„¨oª¥Å¤o Paphiopedilum ‘Delrosi’

œÊ嘵¨ ­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ ­¼˜¦°µ®µ¦ (20 g/l) TDZ (ôM) 2,4-D (ôM) 1 sucrose 2 sucrose 0.45 3 sucrose 4.54 4 sucrose 0.45 4.52 5 sucrose 4.54 4.52 6 sucrose 4.54 13.56 7 glucose

8 glucose 0.45 9 glucose 4.54 10 glucose 0.45 4.52 11 glucose 4.54 4.52

12 glucose 4.54 13.56

13 maltose 14 maltose 0.45 15 maltose 4.54 16 maltose 0.45 4.52 17 maltose 4.54 4.52 18 maltose 4.54 13.56

44

£µ¡š¸É 8 ¨´„¬–³„µ¦Á¡µ³Á¨¸Ê¥Š·Êœ­nªœÄ °Š˜oœ„¨oª¥Å¤o Paphiopedilum ‘Delrosi’

šš¸É 4

Ÿ¨„µ¦š—¨°Š

1. «¹„¬µ­¼˜¦°µ®µ¦š¸ÉÁ®¤µ³­¤˜n°„µ¦´„œÎµ„µ¦Á„·—¥°—šª¸‡¼–‹µ„­ªœ¥°— °Š„¨n oª¥Å¤ o Paphiopedilum ‘Delrosi’ 1.1 Ÿ¨ °Š­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ TDZ, BA, Kn, 2,4-D ¨³ Ads ˜n°„µ¦´„œÎµ„µ¦Á„·—¥°— šª¸‡¼–‹µ„­nªœ¥°— °Š„¨oª¥Å¤ o Paphiopedilum ‘Delrosi’ œÎµ­nªœ¥°— °Š„¨oª¥Å¤o Paphiopedilum ‘Delrosi’ °µ¥» 14 Á—º°œ®¨´Š‹µ„Á¡µ³Á¤¨È— ¤µ Á¡µ³Á¨¸Ê¥Šœ­¼˜¦°µ®µ¦—´—ž¨Š Hyponex [ž»l¥ Hyponex (6.5N-4.5P-19K) ‡ªµ¤Á o¤ oœ 3

„¦´¤˜n°¨·˜¦ + ÁžžÃ˜œ (peptone) ‡ªµ¤Á o¤ oœ 2 „¦´¤˜n°¨·˜¦ + œÊε˜o¤¤´œ ¦´ÉŠ 100 „¦´¤˜n°¨·˜¦] ¦nª¤„´­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ TDZ, BA, Kn, 2,4-D ¨³ Ads ‡ªµ¤Á o¤ oœ˜nµŠÇ —´Š˜µ¦µŠš¸É 1 Áž}œ ¦³¥³Áª¨µ 16 ­´ž—µ®r ¡ªnµ ­¼˜¦°µ®µ¦—´—ž¨Š Hyponex š¸Éž¦³„°—oª¥­µ¦‡ª‡»¤„µ¦Á˜·Ã˜

Kn ‡ªµ¤Á o¤ oœ 4.65 ŤÇ¦Ã¤¨µ¦r ¦nª¤„´ 2,4-D ‡ªµ¤Á o¤ oœ 4.52 ŤÇ¦Ã¤¨µ¦r (­¼˜¦ 12) Ä®o Áž°¦rÁŽÈœ˜r„µ¦Á„·—¥°—ÁŒ¨¸É¥¤µ„š¸É­»— 100 Áž°¦rÁŽÈœ˜r ¨³Ä®o‹Îµœªœ¥°—Ä®¤nÁŒ¨¸É¥¤µ„¨³

­¤ÉεÁ­¤°š¸É­»— 1.4 ¥°— ¦°Š¨Š¤µ‡º°­¼˜¦°µ®µ¦— ´—ž¨Š Hyponex š¸Éž¦³„°—ª¥­µ¦‡ª‡o »¤„µ¦ Á˜·Ã˜ TDZ ‡ªµ¤Á o¤ oœ 0.45 ŤÇ¦Ã¤¨µ¦r (­¼˜¦ 2) ®¦º° BA ‡ªµ¤Á o¤ oœ 4.44 ŤÇ¦Ã¤¨µ¦r ¦nª¤„´ 2,4-D ‡ªµ¤Á o¤ oœ 4.52 ŤÇ¦Ã¤¨µ¦r (­¼˜¦ 11) ®¦º° TDZ ‡ªµ¤Á o¤ oœ 0.45 ŤÇ¦Ã¤ ¨µ¦r ¦nª¤„´ 2,4-D ‡ªµ¤Á o¤ oœ 4.52 ŤÇ¦Ã¤¨µ¦r (­¼˜¦ 8) Ä®oÁž°¦rÁŽÈœ˜r„µ¦Á„·—¥°—Ä®¤nÁŒ¨¸É¥ 90 Áž°¦rÁŽÈœ˜r ¨³¤¸‹Îµœªœ¥°—Ä®¤nÁŒ¨¸É¥ 1.6 1.4 ¨³ 1.2 ¥°— ˜µ¤¨Îµ—´ (˜µ¦µŠš¸É 8, ¦¼žš¸É 9D, 9A, 9C ¨³ 9B) Ĝ –³š¸É­¼˜¦°µ®µ¦°ºÉœÇ Ä®oÁž°¦ÁŽr Ȝ˜r„µ¦Á„—¥°—Ä®¤· nÁŒ¨¸É¥¦³®ªnµŠ 30-70 Áž°¦rÁŽÈœ˜r ¨³¤¸‹Îµœªœ¥°—Ä®¤nÁŒ¨¸É¥¦³®ªnµŠ 0.3-1.3 ¥°— ­nªœœÊ宜´„­— ‹ÎµœªœÄ ¨³‹Îµœªœ ¦µ„ÁŒ¨¸É¥¥°—Ä®¤n °Šš»„­¼˜¦°µ®µ¦ ¤¸‡nµÁŒ¨¸É¥Å¤n˜„˜nµŠ„´œ°¥nµŠ¤¸œ´¥­Îµ‡´šµŠ­™·˜·š¸ÉnªŠ‡ªµ¤ ÁºÉ°¤´Éœ 95 Áž°¦rÁŽÈœ˜r ­Îµ®¦´œÊ宜„­—¦ª¤Â¨³œ´ Ê宜„­—¥°—ÁŒ¨´ ¸É¥ °Š¥°—Á¦·É¤˜oœ ¡ªnµ ­¼˜¦°µ®µ¦ —´—ž¨Š Hyponex š¸Éž¦³„°—oª¥­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ TDZ ‡ªµ¤Á o¤ oœ 0.45 ŤÇ¦Ã¤¨µ¦r ¦nª¤„´ 2,4-D ‡ªµ¤Á o¤ oœ 4.52 ŤÇ¦Ã¤¨µ¦r (­¼˜¦ 8) Ä®oœÊ宜„­—¦ª¤Â¨´ ³œÊ宜„­—¥°—´ ÁŒ¨¸É¥¤µ„š¸É­»— 977.1 ¨³ 649.1 ¤·¨¨·„¦´¤ ˜µ¤¨Îµ—´ (˜µ¦µŠš¸É 9, ¦¼žš¸É 9B) ­nªœœÊ宜„­—¦µ„´ 45 46

¨³‹Îµœªœ¦µ„ÁŒ¨¸É¥ ¡ªnµ ­¼˜¦°µ®µ¦—´—ž¨Š Hyponex š¸Éž¦³„°—oª¥­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ TDZ ‡ªµ¤Á ¤ o oœ 4.54 ŤÇ¦Ã¤¨µ¦r ¦nª¤„´ 2,4-D ‡ªµ¤Á o¤ oœ 4.52 ŤÇ¦Ã¤¨µ¦r (­¼˜¦ 9) Ä®o œÊ宜„­—¦µ„´ ¨³‹Îµœªœ¦µ„ÁŒ¨¸É¥¤µ„š¸É­»— 402.3 ¤·¨¨·„¦´¤ ¨³ 4.9 ¦µ„ ˜µ¤¨µ—Î ´ (˜µ¦µŠš ¸É 9, ¦¼žš¸É 9F) ¨³¡ªnµ š»„­¼˜¦°µ®µ¦š¸Éž¦³„°—oª¥ Ads ‡ªµ¤Á o¤ oœ 27.1 ŤÇ¦Ã¤¨µ¦r šÎµÄ®o¦µ„ ¤¸¨´„¬–³Á¨È„ ­´Êœ ¤¸ œ¦µ„œo°¥®¦º°Å¤n¤¸ œ¦µ„ (¦¼žš¸É 9E) Ĝ –³š¸É­¼˜¦°µ®µ¦—´—ž¨Š Hyponex š¸Éž¦³„°—ª¥­µ¦‡ª‡o »¤„µ¦Á˜·Ã˜ Kn ‡ªµ¤Á o¤ oœ 4.65 ŤÇ¦Ã¤¨µ¦r ¦nª¤„´ 2,4-D ‡ªµ¤ Á o¤ oœ 4.52 ŤÇ¦Ã¤¨µ¦r (­¼˜¦ 12) Ä®o‹ÎµœªœÄÁŒ¨¸É¥¤µ„š¸É­»— 6.0 ĝ (˜µ¦µŠš¸É 9, ¦¼žš¸É 9D)

47

˜µ¦µŠš¸É 8 Ÿ¨ °Š­¼˜¦°µ®µ¦—´—ž¨Š Hyponex ¦nª¤„´­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ TDZ, BA, Kn, 2,4-D ¨³ Ads ˜n°„µ¦´„œÎµ„µ¦Á„·—¥°—šª¸‡¼–‹µ„­ªœ¥°—n °Š„¨oª¥Å¤ o Paphiopedilum ‘Delrosi’ ®¨´Š‹µ„„µ¦Á¡µ³Á¨¸Ê¥ŠÁžœ¦³¥³Áª¨µ} 16 ­´ž—µ® r ­¼˜¦ ­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ (ôM) „µ¦Á‹¦·Á˜·Ã˜ °Š¥°—Ä®¤n1/ Hyponex TDZ BA Kn 2,4-D Ads „µ¦Á„·—¥°—3/ (%) ‹Îµœªœ¥°—3/ œÊ宜´„­—¦ª¤2/,4/ (mg) ‹ÎµœªœÄ3/ ‹Îµœªœ¦µ„3/ 1 30.00±15.28 c 0.30±0.15 d 25.17±7.80 a 2.0±0.0 a 0.0±0.0 a 2 0.45 90.00±10.00 ab 1.60±0.52 ab 13.06±4.76 a 1.9±0.1 a 0.2±0.0 a 3 4.54 30.00±15.28 c 0.40±0.22 d 27.53±17.30 a 2.2±0.4 a 1.0±0.0 a 4 2.22 50.00±16.67 abc 0.60±0.22 bcd 29.57±6.66 a 2.4±0.2 a 0.8±0.3 a

5 4.44 70.00±15.28 abc 1.30±0.40 abcd 14.20±2.08 a 2.1±0.2 a 0.3±0.0 a 6 4.65 60.00±16.33 abc 0.60±0.16 bcd 25.80±12.07 a 2.2±0.3 a 1.5±0.5 a 7 9.29 40.00±16.33 bc 0.40±0.16 cd 30.85±7.70 a 2.5±0.3 a 1.0±0.0 a

8 0.45 4.52 90.00±10.00 ab 1.20±0.20 abc 43.15±16.41 a 2.5±0.2 a 1.2±0.0 a 9 4.54 4.52 30.00±15.28 c 0.40±0.22 d 28.00±11.32 a 2.2±0.2 a 0.0±0.0 a

10 2.22 4.52 70.00±15.28 abc 0.80±0.20 abcd 47.34±15.32 a 2.6±0.2 a 1.5±0.8 a 11 4.44 4.52 90.00±10.00 ab 1.40±0.27 ab 18.70±4.55 a 2.0±0.2 a 0.8±0.3 a 12 4.65 4.52 100.00±0.00 a 1.40±0.22 a 24.03±9.46 a 2.2±0.3 a 1.1±0.1 a 13 0.45 27.1 30.00±15.28 c 0.30±0.15 d 16.17±10.36 a 2.3±0.3 a 0.0±0.0 a 14 4.54 27.1 40.00±16.33 bc 0.50±0.22 cd 20.98±4.45 a 2.4±0.2 a 0.8±0.3 a 15 2.22 27.1 70.00±15.28 abc 0.90±0.23 abcd 28.76±7.93 a 2.5±0.2 a 1.3±0.6 a 16 4.44 27.1 30.00±15.28 c 0.30±0.15 d 38.27±17.94 a 2.3±0.7 a 1.0±0.0 a 1/˜´ªÁ¨ (‡nµÁŒ¨¸É¥ ± S.E.) š¸É˜µ¤—oª¥˜´ª°´„¬¦Á®¤º°œ„´œÄœÂœª—·ÉŠ ­—ŠªnµÅ¤n¤¸‡ªµ¤Â˜„˜nµŠšµŠ­™·˜·š¸É¦³—´‡ªµ¤ÁºÉ°¤´Éœ 95 Áž°¦rÁŽÈœ˜r (P = 0.05) šÎµ„µ¦š—¨°Š 10 ŽÊε; 2/×¥ª·›¸„µ¦Áž¦¸¥Áš¥Â¸ DMRT; 3/×¥ª·›¸„µ¦Áž¦¸¥Áš¸¥Â Kruskal-Wallis H ¨³ Mann-Whitney U; 4/œÊ宜´„­—¦ª¤ = œÊ宜´„­—˜oœ¦ª¤„´œÊ宜´„­—¦µ„ 48

˜µ¦µŠš¸É 9 Ÿ¨ °Š­¼˜¦°µ®µ¦—´—ž¨Š Hyponex ¦nª¤„´­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ TDZ, BA, Kn, 2,4-D ¨³ Ads ˜n°„µ¦Á‹¦·Á˜·Ã˜ °Š­nªœ¥°—Á¦·É¤˜oœ °Š „¨oª¥Å¤ o Paphiopedilum ‘Delrosi’ ®¨´Š‹µ„„µ¦Á¡µ³Á¨¥ŠÁž¸Ê }œ¦³¥³Áª¨µ 16 ­´ž—µ® r ­¼˜¦ ­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ (ôM) „µ¦Á‹¦·Á˜·Ã˜ °Š¥°—Á¦·É¤˜oœ1/ Hyponex TDZ BA Kn 2,4-D Ads œÊ宜´„­—¦ª¤2/,4/ (mg) œÊ宜´„­—¥°—2/ (mg) œÊ宜´„­—¦µ„2/ (mg) ‹ÎµœªœÄ3/ ‹Îµœªœ¦µ„2/ 1 819.20±55.81 abc 516.40±29.50 bc 302.80±37.26 bc 5.1±0.2 d 4.4±0.3 ab 2 0.45 891.60±57.99 ab 572.50±35.73 abc 319.10±36.02 abc 5.4±0.2 bcd 4.8±0.2 a 3 4.54 873.20±48.46 ab 521.80±29.99 bc 351.40±22.79 ab 5.2±0.2 cd 4.5±0.2 ab 4 2.22 848.50±47.06 abc 578.10±34.54 abc 270.40±19.85 bc 5.4±0.2 bcd 3.9±0.2 bcd

5 4.44 843.20±73.75 abc 527.40±45.92 bc 315.80±32.86 abc 5.4±0.3 abcd 3.9±0.2 bcd 6 4.65 898.00±44.62 ab 580.50±30.78 abc 317.50±20.78 abc 5.6±0.2 abcd 4.1±0.2 abc 7 9.29 821.13±67.27 abc 592.10±46.03 abc 229.03±28.89 cd 5.6±0.2 abcd 3.8±0.2 bcd

8 0.45 4.52 977.10±42.71 a 649.10±34.04 a 328.00±16.71 ab 5.8±0.1 abc 4.3±0.3 ab 9 4.54 4.52 962.30±55.06 ab 560.00±31.75 abc 402.30±27.86 a 5.5±0.2 abcd 4.9±0.3 a

10 2.22 4.52 786.16±38.78 bcd 515.00±18.98 bc 271.16±26.03 bc 5.9±0.1 ab 4.6±0.3 ab 11 4.44 4.52 833.00±47.32 abc 569.70±30.21 abc 263.30±26.83 bc 5.7±0.2 abcd 4.6±0.3 ab 12 4.65 4.52 898.24±54.68 ab 612.80±29.07 ab 285.44±36.17 bc 6.0±0.1 a 4.3±0.3 ab 13 0.45 27.1 680.14±34.40 cd 524.70±31.09 bc 155.44±12.01 d 5.7±0.2 abc 3.2±0.2 d 14 4.54 27.1 642.24±54.07 d 474.70±43.20 c 167.54±18.89 d 5.4±0.2 bcd 3.3±0.2 cd 15 2.22 27.1 918.90±69.03 ab 622.50±38.15 ab 296.40±37.98 bc 5.9±0.1 ab 4.6±0.3 ab 16 4.44 27.1 934.70±71.44 ab 633.40±53.88 ab 301.30±28.29 bc 5.4±0.2 bcd 4.9±0.3 a 1/˜´ªÁ¨ (‡nµÁŒ¨¸É¥ ± S.E.) š¸É˜µ¤—oª¥˜´ª°´„¬¦Á®¤º°œ„´œÄœÂœª—·ÉŠ ­—ŠªnµÅ¤n¤¸‡ªµ¤Â˜„˜nµŠšµŠ­™·˜·š¸É¦³—´‡ªµ¤ÁºÉ°¤´Éœ 95 Áž°¦rÁŽÈœ˜r (P = 0.05) šÎµ„µ¦š—¨°Š 10 ŽÊε; 2/×¥ª·›¸„µ¦Áž¦¸¥Áš¥Â¸ DMRT; 3/×¥ª·›¸„µ¦Áž¦¸¥Áš¸¥Â Kruskal-Wallis H ¨³ Mann-Whitney U; 4/œÊ宜´„­—¦ª¤ = œÊ宜´„­—˜oœ¦ª¤„´œÊ宜´„­—¦µ„ 49

£µ¡š¸É 9 Ÿ¨ °Š­¼˜¦°µ®µ¦—´—ž¨Š Hyponex ¦nª¤„´­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ TDZ, BA, Kn, 2,4-D ¨³ Ads ˜n° „µ¦Á‹¦·Á˜·Ã˜Â¨³„µ¦´„œÎµ„µ¦Á„·—¥°—šª¸‡¼–‹µ„­nªœ¥°— °Š„¨oª¥Å¤o Paphiopedilum ‘Delrosi’ ®¨´Š‹µ„„µ¦Á¡µ³Á¨¸Ê¥ŠÁž}œ¦³¥³Áª¨µ 16 ­´ž—µ®r A = ­¼˜¦°µ®µ¦—´—ž¨Š Hyponex + TDZ 0.45 ôM (­¼˜¦ 2) B = ­¼˜¦°µ®µ¦—´—ž¨Š Hyponex + TDZ 0.45 ôM + 2,4-D 4.52 ôM (­¼˜¦ 8) C = ­¼˜¦°µ®µ¦—´—ž¨Š Hyponex + BA 4.44 ôM + 2,4-D 4.52 ôM (­¼˜¦ 11)

50

£µ¡š¸É 9 (˜n°) Ÿ¨ °Š­¼˜¦°µ®µ¦—´—ž¨Š Hyponex ¦nª¤„´­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ TDZ, BA, Kn, 2,4-D ¨³ Ads ˜n°„µ¦Á‹¦·Á˜·Ã˜Â¨³„µ¦´„œÎµ„µ¦Á„·—¥°—šª¸‡¼–‹µ„­nªœ¥°— °Š„¨oª¥Å¤o Paphiopedilum ‘Delrosi’ ®¨´Š‹µ„„µ¦Á¡µ³Á¨¸Ê¥ŠÁž}œ¦³¥³Áª¨µ 16 ­´ž—µ®r D = ­¼˜¦°µ®µ¦—´—ž¨Š Hyponex + Kn 4.65 ôM + 2,4-D 4.52 ôM (­¼˜¦ 12) E = ­¼˜¦°µ®µ¦—´—ž¨Š Hyponex + TDZ 0.45 ôM + Ads 27.1 ôM (­¼˜¦ 13) F = ­¼˜¦°µ®µ¦—´—ž¨Š Hyponex + TDZ 4.54 ôM + 2,4-D 4.52 ôM (­¼˜¦ 9)

51

1.2 «¹„¬µŸ¨ °Š­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ TDZ, BA, Kn ¨³ 2,4-D ˜n°„µ¦´„œÎµ„µ¦Á„·—¥°—šª¸‡¼– ‹µ„­nªœ¥°— °Š„¨oª¥Å¤ o Paphiopedilum ‘Delrosi’ œÎµ­nªœ¥°— °Š„¨oª¥Å¤o Paphiopedilum ‘Delrosi’ °µ¥» 19 Á—º°œ®¨´Š‹µ„Á¡µ³Á¤¨È— ¤µ Á¡µ³Á¨¸Ê¥Šœ­¼˜¦°µ®µ¦—´—ž¨Š Hyponex [ž»l¥ Hyponex (6.5N-4.5P-19K) ‡ªµ¤Á o¤ oœ 3 „¦´¤˜n°¨·˜¦ + ÁžžÃ˜œ (peptone) ‡ªµ¤Á o¤ oœ 2 „¦´¤˜n°¨·˜¦ + œÊε˜o¤¤´œ ¦´ÉŠ 100 „¦´¤˜n°¨·˜¦] ¦nª¤„´­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ TDZ, BA, Kn ¨³ 2,4-D ‡ªµ¤Á o¤ oœ˜nµŠÇ —´Š˜µ¦µŠš¸É 2 Áž}œ ¦³¥³Áª¨µ 16 ­´ž—µ®r ¡ªnµ ­¼˜¦°µ®µ¦—´—ž¨Š Hyponex š¸Éž¦³„°—oª¥­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ TDZ ‡ªµ¤Á ¤ o oœ 0.45 ŤÇ¦Ã¤¨µ¦r ¦nª¤„´ 2,4-D ‡ªµ¤Á o¤ oœ 4.52 ŤÇ¦Ã¤¨µ¦r (­¼˜¦ 8) Ä®o Áž°¦rÁŽÈœ˜r„µ¦Á„·—¥°—Ä®¤nÁŒ¨¸É¥¤µ„š¸É­»— 90 Áž°¦rÁŽÈœ˜r ¨³Ä®o‹Îµœªœ¥°—Ä®¤nÁŒ¨¸É¥¤µ„¨³ ­¤ÉεÁ­¤°š¸É­»— 1.3 ¥°— ¦°Š¨Š¤µ‡º°­¼˜¦°µ®µ¦—´—ž¨Š Hyponex š¸Éž¦³„°—ª¥­µ¦‡ª‡o »¤„µ¦ Á˜·Ã˜ TDZ ‡ªµ¤Á o¤ oœ 0.45 ŤÇ¦Ã¤¨µ¦r (­¼˜¦ 2) ®¦º° BA ‡ªµ¤Á o¤ oœ 8.88 ŤÇ¦Ã¤¨µ¦r

¦nª¤„´ 2,4-D ‡ªµ¤Á o¤ oœ 13.56 ŤÇ¦Ã¤¨µ¦r (­¼˜¦ 13) Ä®oÁž°¦rÁŽÈœ˜r„µ¦Á„·—¥°—Ä®¤nÁŒ¨¸É¥ 80 Áž°¦rÁŽÈœ˜r ¨³¤¸‹Îµœªœ¥°—Ä®¤nÁŒ¨¸É¥ 1.5 ¨³ 1.1 ¥°— ˜µ¤¨Îµ—´ (˜µ¦µŠš¸É 10, ¦¼žš¸É 10B, 10A

¨³ 10D) Ĝ –³š¸É­¼˜¦°µ®µ¦°ºÉœÇ Ä®oÁž°¦rÁŽÈœ˜r„µ¦Á„·—¥°—Ä®¤nÁŒ¨¸É¥¦³®ªµŠn 10-60 Áž°¦rÁŽÈœ˜r ¨³¤¸‹Îµœªœ¥°—Ä®¤nÁŒ¨¸É¥¦³®ªnµŠ 0.3-1.0 ¥°— Ĝ—µœœo Ê宜´„­—¥°—Ä®¤nÁŒ¨¸É¥ ¡ªnµ ­¼˜¦

°µ®µ¦—´—ž¨Š Hyponex š¸Éž¦³„°—ª¥­µ¦‡ª‡o »¤„µ¦Á˜·Ã˜ BA ‡ªµ¤Á o¤ oœ 8.88 ŤÇ¦Ã¤

¨µ¦r ¦nª¤„´ 2,4-D ‡ªµ¤Á ¤ o oœ 13.56 ŤÇ¦Ã¤¨µ¦r (­¼˜¦ 13) Ä®oœÊ宜„­—¥°—Ä®¤´ nÁŒ¨¥¤µ„¸É š¸É­»— 316.7 ¤·¨¨·„¦´¤ (˜µ¦µŠš¸É 10, ¦¼žš¸É 10C ) ­nªœ‹ÎµœªœÄ ¨³‹Îµœªœ¦µ„¥°—Ä®¤nÁŒ¨¸É¥ °Šš»„ ­¼˜¦°µ®µ¦¤¸‡nµÁŒ¨¸É¥Å¤n˜„˜nµŠ„´œ°¥nµŠ¤¸œ´¥­Îµ‡´šµŠ­™·˜·š¸ÉnªŠ‡ªµ¤ÁºÉ°¤´Éœ 95 Áž°¦rÁŽÈœ˜r ­Îµ®¦´œÊ宜„­—¦ª¤´ œÊ宜´„­—¥°— œÊ宜„­—¦µ„´ ‹ÎµœªœÄ ¨³‹Îµœªœ¦µ„ ÁŒ¨¸É¥ °Š¥°—Á¦·É¤˜oœ ¡ªnµ ­nªœÄ®n¤¸‡nµÁŒ¨¸É¥¤µ„ ¹ÊœÁ¤°Á¡µ³Á¨ºÉ ¸Ê¥Šœ­¼˜¦°µ®µ¦š¸Éž¦³„° —oª¥ ­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ 2,4-D š´ÊŠ­°Š‡ªµ¤Á o¤ oœ ‡º° 4.52 ¨³ 13.56 ŤÇ¦Ã¤¨µ¦r (˜µ¦µŠš¸É 11)

52

˜µ¦µŠš¸É 10 Ÿ¨ °Š­¼˜¦°µ®µ¦—´—ž¨Š Hyponex ¦nª¤„­µ¦‡ª‡´ »¤„µ¦Á˜·Ã˜ TDZ, BA, Kn ¨³ 2,4-D ˜n°„µ¦´„œÎµ„µ¦Á„—¥°—šª· ¸‡¼–‹µ„­nªœ¥°— °Š „¨oª¥Å¤ o Paphiopedilum ‘Delrosi’ ®¨´Š‹µ„„µ¦Á¡µ³Á¨¥ŠÁž¸Ê }œ¦³¥³Áª¨µ 16 ­´ž—µ® r ­¼˜¦ ­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ (ôM) „µ¦Á‹¦·Á˜·Ã˜ °Š¥°—Ä®¤n1/ Hyponex TDZ BA Kn 2,4-D „µ¦Á„·—¥°—3/ (%) ‹Îµœªœ¥°—3/ œÊ宜´„­—¦ª¤3/,4/ (mg) ‹ÎµœªœÄ3/ ‹Îµœªœ¦µ„3/ 1 30.00±15.28 bc 0.30±0.15 c 52.67±25.17 abc 2.7±0.3 a 0.7±0.3 a 2 0.45 80.00±13.33 ab 1.50±0.45 ab 116.13±70.41 abc 2.7±0.3 a 0.7±0.4 a

3 2.25 50.00±16.67 abc 0.60±0.22 abc 19.80±6.08 c 2.0±0.0 a 0.2±0.2 a 4 4.44 60.00±16.33 abc 0.80±0.25 abc 106.67±49.97 abc 2.6±0.3 a 0.8±0.5 a 5 8.88 40.00±16.33 abc 1.00±0.47 abc 143.00±45.80 ab 2.8±0.4 a 1.0±0.4 a 6 2.32 40.00±16.33 abc 0.70±0.30 abc 121.00±43.03 ab 3.0±0.2 a 1.5±0.2 a 7 4.65 30.00±15.28 bc 0.50±0.27 bc 201.67±104.83 ab 2.7±0.4 a 1.2±0.7 a

8 0.45 4.52 90.00±10.00 a 1.30±0.21 a 187.89±46.32 ab 2.8±0.3 a 1.4±0.3 a 9 0.45 13.56 60.00±16.33 abc 0.90±0.38 abc 90.83±66.01 bc 2.4±0.2 a 0.5±0.4 a 10 2.25 4.52 50.00±16.67 abc 0.60±0.22 abc 190.20±72.73 ab 2.9±0.5 a 1.4±0.5 a

11 2.25 13.56 10.00±10.00 c 0.10±0.10 c 141.00±0.00 ab 3.0±0.0 a 2.0±0.0 a 12 4.44 13.56 30.00±15.28 bc 0.30±0.15 c 0.24±0.11 d 3.3±0.3 a 3.3±0.9 a 13 8.88 13.56 80.00±13.33 ab 1.10±0.23 ab 316.71±85.42 a 3.0±0.3 a 1.6±0.4 a 1/˜´ªÁ¨ (‡nµÁŒ¨¸É¥ ± S.E.) š¸É˜µ¤—oª¥˜´ª°´„¬¦Á®¤º°œ„´œÄœÂœª—·ÉŠ ­—ŠªnµÅ¤n¤¸‡ªµ¤Â˜„˜nµŠšµŠ­™·˜·š¸É¦³—´‡ªµ¤ÁºÉ°¤´Éœ 95 Áž°¦rÁŽÈœ˜r (P = 0.05) šÎµ„µ¦š—¨°Š 10 ŽÊε; 2/×¥ª·›¸„µ¦Áž¦¸¥Áš¥Â¸ DMRT; 3/×¥ª·›¸„µ¦Áž¦¸¥Áš¸¥Â Kruskal-Wallis H ¨³ Mann-Whitney U; 4/œÊ宜´„­—¦ª¤ = œÊ宜´„­—˜oœ¦ª¤„´œÊ宜´„­—¦µ„

53

˜µ¦µŠš¸É 11 Ÿ¨ °Š­¼˜¦°µ®µ¦—´—ž¨Š Hyponex ¦nª¤„­µ¦‡ª‡´ »¤„µ¦Á˜·Ã˜ TDZ, BA, Kn ¨³ 2,4-D ˜n°„µ¦Á‹¦·Á˜·Ã˜ °Š¥°—Á¦·É¤˜œ °Š„¨o oª¥Å¤ o Paphiopedilum ‘Delrosi’ ®¨´Š‹µ„„µ¦Á¡µ³Á¨¸Ê¥ŠÁžœ¦³¥³Áª¨µ} 16 ­´ž—µ® r ­¼˜¦ ­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ (ôM) „µ¦Á‹¦·Á˜·Ã˜ °Š¥°—Á¦·É¤˜oœ1/ Hyponex TDZ BA Kn 2,4-D œÊ宜´„­—¦ª¤2/,4/ (mg) œÊ宜´„­—¥°—2/ (mg) œÊ宜´„­—¦µ„2/ (mg) ‹ÎµœªœÄ3/ ‹Îµœªœ¦µ„2/ 1 1502.20±98.39 e 855.20±43.50 d 647.00±59.29 d 6.0±0.0 c 5.7±0.2 e 2 0.45 1942.50±141.03 cde 936.10±64.20 cd 1006.40±86.81 abc 6.5±0.2 abc 6.8±0.2 bcde

3 2.25 1805.80±83.67 cde 1073.70±37.87 bcd 732.10±76.40 cd 6.5±0.2 abc 6.7±0.3 cde 4 4.44 1852.40±202.22 cde 1034.70±76.73 cd 817.70±144.76 bcd 6.3±0.2 bc 7.0±0.4 bcd 5 8.88 1472.20±89.37 e 910.00±68.78 cd 562.20±34.87 d 6.6±0.2 ab 6.7±0.4 cde 6 2.32 1616.90±122.27 e 964.10±86.48 cd 652.80±67.69 d 6.3±0.2 bc 6.4±0.3 cde 7 4.65 1754.40±160.76 de 1013.00±80.93 cd 741.40±110.94 cd 6.7±0.2 ab 6.2±0.4 de

8 0.45 4.52 2129.30±195.09 bcd 1149.90±101.85 bc 979.40±102.13 abc 7.2±0.2 a 7.3±0.4 abcd 9 0.45 13.56 2571.10±122.80 ab 1572.90±74.63 a 998.20±57.04 abc 6.7±0.2 ab 7.2±0.2 abcd 10 2.25 4.52 2276.40±139.18 abc 1295.90±75.02 b 980.50±76.21 abc 6.9±0.2 ab 7.3±0.3 abcd

11 2.25 13.56 2726.40±154.27 a 1538.50±66.53 a 1187.90±104.79 a 6.5±0.2 abc 7.9±0.3 ab 12 4.44 13.56 2742.90±253.70 a 1647.80±110.69 a 1095.10±149.93 ab 7.0±0.2 ab 7.4±0.5 abc 13 8.88 13.56 2714.70±195.21 a 1553.70±122.25 a 1161.00±94.35 a 6.6±0.2 ab 8.3±0.5 a 1/˜´ªÁ¨ (‡nµÁŒ¨¸É¥ ± S.E.) š¸É˜µ¤—oª¥˜´ª°´„¬¦Á®¤º°œ„´œÄœÂœª—·ÉŠ ­—ŠªnµÅ¤n¤¸‡ªµ¤Â˜„˜nµŠšµŠ­™·˜·š¸É¦³—´‡ªµ¤ÁºÉ°¤´Éœ 95 Áž°¦rÁŽÈœ˜r (P = 0.05) šÎµ„µ¦š—¨°Š 10 ŽÊε; 2/×¥ª·›¸„µ¦Áž¦¸¥Áš¥Â¸ DMRT; 3/×¥ª·›¸„µ¦Áž¦¸¥Áš¸¥Â Kruskal-Wallis H ¨³ Mann-Whitney U; 4/œÊ宜´„­—¦ª¤ = œÊ宜´„­—˜oœ¦ª¤„´œÊ宜´„­—¦µ„

54

£µ¡š¸É 10 Ÿ¨ °Š­¼˜¦°µ®µ¦—´—ž¨Š Hyponex ¦nª¤„´­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ TDZ, BA, Kn ¨³ 2,4-D ˜n°„µ¦ Á‹¦·Á˜·Ã˜Â¨³„µ¦´„œÎµ„µ¦Á„·—¥°—šª¸‡¼–‹µ„­nªœ¥°— °Š„¨oª¥Å¤o Paphiopedilum ‘Delrosi’ ®¨´Š‹µ„„µ¦Á¡µ³Á¨¸Ê¥ŠÁž}œ¦³¥³Áª¨µ 16 ­´ž—µ®r A = ­¼˜¦°µ®µ¦—´—ž¨Š Hyponex + TDZ 0.45 ôM (­¼˜¦ 2) B = ­¼˜¦°µ®µ¦—´—ž¨Š Hyponex + TDZ 0.45 ôM + 2,4-D 4.52 ôM (­¼˜¦ 8) C = ­¼˜¦°µ®µ¦—´—ž¨Š Hyponex + BA 8.88 ôM + 2,4-D 13.56 ôM (­¼˜¦ 13) 55

2. «¹„¬µœ·— °ŠœÊ嘵¨š¸ÉÁ®¤µ³­¤˜n°„µ¦´„œÎµ„µ¦Á„—¥°—šª· ¸‡¼–‹µ„­nªœ¥°— °Š„¨oª¥Å¤ o Paphiopedilum ‘Delrosi’ œÎµ­nªœ¥°— °Š„¨oª¥Å¤ o Paphiopedilum ‘Delrosi’ ¤µÁ¡µ³Á¨¸Ê¥Šœ­¼˜¦°µ®µ¦ —´—ž¨Š Hyponex [ž»l¥ Hyponex (6.5N-4.5P-19K) ‡ªµ¤Á o¤ oœ 3 „¦´¤˜n°¨·˜¦ + ÁžžÃ˜œ (peptone) ‡ªµ¤Á o¤ oœ 2 „¦´¤˜n°¨·˜¦ + œÊε˜o¤¤´œ ¦´ÉŠ 100 „¦´¤˜n°¨·˜¦] š¸Éž¦³„°—oª¥œÊ嘵¨ 3 œ·— ‡º° Ž¼Ã‡¦­ „¨¼Ã‡­ ¨³¤°¨Ã˜­ ‡ªµ¤Á o¤ oœ 20 „¦´¤˜n°¨·˜¦ ¦nª¤„´­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ TDZ ‡ªµ¤Á ¤ o oœ 0.45 ŤÇ¦Ã¤¨µ¦r ¦nª¤„´ 2,4-D ‡ªµ¤Á o¤ oœ 4.52 ŤÇ¦Ã¤¨µ¦r ®¦º° Kn ‡ªµ¤Á o¤ oœ 4.65 ŤÇ¦Ã¤¨µ¦r ¦nª¤„´ 2,4-D ‡ªµ¤Á ¤ o oœ 4.52 ŤÇ¦Ã¤¨µ¦r —´Š˜µ¦µŠš¸É 3 Áž}œ ¦³¥³Áª¨µ 16 ­´ž—µ®r ¡ªnµ š»„­¼˜¦°µ®µ¦Ä®oÁž°¦rÁŽÈœ˜r„µ¦Á„·—¥°—Ä®¤nÁŒ¨¸É¥Å¤Â˜„˜n nµŠ„´œ°¥nµŠ ¤¸œ´¥­Îµ‡´šµŠ­™·˜·š¸ÉnªŠ‡ªµ¤ÁºÉ°¤´Éœ 90 Áž°¦rÁŽÈœ˜r °¥µŠÅ¦„n Șµ¤ ­¼˜¦°µ®µ¦—´—ž¨Š Hyponex š¸Éž¦³„°—ª¥œo Ê嘵¨„¨¼Ã‡­ (­¼˜¦ 3) ®¦°Žº ¼Ã‡¦­ (­¼˜¦ 1) ‡ªµ¤Á o¤ oœ 20 „¦´¤˜n°¨·˜¦ ¨³­µ¦

‡ª‡»¤„µ¦Á˜Ã˜· TDZ ‡ªµ¤Á o¤ oœ 0.45 ŤÇ¦Ã¤¨µ¦r ¦nª¤„´ 2,4-D ‡ªµ¤Á o¤ oœ 4.52 ŤÇ¦ 䨵¦r Ä®oÁž°¦rÁŽÈœ˜r„µ¦Á„—¥°—Ä®¤· nÁŒ¨¥¤µ„š¸É ¸É­»— 80 Áž°¦rÁŽÈœ˜r ¨³Ä®o‹Îµœªœ¥°—Ä®¤nÁŒ¨¸É¥

1.8 ¨³ 1.5 ¥°— ˜µ¤¨Îµ—´ (˜µ¦µŠš ¸É 12, ¦¼žš¸É 11B ¨³ 11A) Ĝ –³š¸É­¼˜¦°µ®µ¦°ºÉœÇ Ä®o Áž°¦rÁŽÈœ˜r„µ¦Á„·—¥°—Ä®¤nÁŒ¨¸É¥¦³®ªµŠn 50-70 Áž°¦rÁŽÈœ˜r ¨³Ä®o‹Îµœªœ¥°—Ä®¤nÁŒ¨¸É¥¦³®ªnµŠ

0.5-1.6 ¥°— ­Îµ®¦´œÊ宜´„­— ¨³‹Îµœªœ¦µ„ÁŒ¨¸É¥ °Š¥°—Ä®¤ n ¡ªnµ ­¼˜¦°µ®µ¦—´—ž¨Š

Hyponex š¸Éž¦³„°—ª¥œo Ê嘵¨Ž¼Ã‡¦­ ‡ªµ¤Á o¤ oœ 20 „¦´¤˜n°¨·˜¦ ¨³­µ¦‡ª‡»¤„µ¦Á‹¦· Á˜·Ã˜ Kn ‡ªµ¤Á o¤ oœ 4.65 ŤÇ¦Ã¤¨µ¦ r ¦nª¤„´ 2,4-D ‡ªµ¤Á o¤ oœ 4.52 ŤÇ¦Ã¤¨µ¦r (­¼˜¦ 2) Ä®¥°—Ä®¤o nš¸É¤¸œÊ宜„­—´ ¨³‹Îµœªœ¦µ„ÁŒ¨¸É¥¤µ„š¸É­»— 252.43 ¤·¨¨·„¦´¤ ¨³ 1.4 ¦µ„ ˜µ¤¨Îµ—´ (˜µ¦µŠš¸É 12, ¦¼žš¸É 11C) ­nªœ‹ÎµœªœÄ¥°—Ä®¤nÁŒ¨¸É¥ ¡ªnµ š»„­¼˜¦°µ®µ¦Ä®o‹ÎµœªœÄ ¥°—Ä®¤nÁŒ¨¸É¥Å¤n˜„˜nµŠ„´œ°¥nµŠ¤¸œ´¥­Îµ‡šµŠ­™´ ·˜·š¸ÉnªŠ‡ªµ¤ÁºÉ°¤´Éœ 90 Áž°¦rÁŽÈœ˜r ­Îµ®¦´œÊ宜„­—¦ª¤´ œÊ宜´„­—¥°— œÊ宜„­—¦µ„´ ¨³‹Îµœªœ¦µ„ÁŒ¨¸É¥ °Š¥°— Á¦·É¤˜oœ­nªœÄ®n ¡ªnµ ­¼˜¦°µ®µ¦—´—ž¨Š Hyponex š¸Éž¦³„°—ª¥œo Ê嘵¨Ž¼Ã‡¦­ ®¦º°„¨¼Ã‡­ ‡ªµ¤Á o¤ oœ 20 „¦´¤˜n°¨·˜¦ ¦nª¤„´­µ¦‡ª‡»¤„µ¦Á˜Ã˜š· ´ÊŠ 2  (­¼˜¦ 1-4) Ä®o‡nµÁŒ¨¸É¥ ¤µ„„ªnµ­¼˜¦°µ®µ¦—´—ž¨Š Hyponex š¸Éž¦³„°—ª¥œo Ê嘵¨¤°¨Ã˜­°¥nµŠ¤¸œ´¥­Îµ‡´šµŠ­™·˜·š¸É nªŠ‡ªµ¤ÁºÉ°¤´Éœ 90 Áž°¦rÁŽÈœ˜r ¥„Áªœœo Ê宜´„­—¥°— °Š¥°—Á¦¤˜·É oœš¸ÉÁ¡µ³Á¨¸Ê¥Šœ­¼˜¦°µ®µ¦ —´—ž¨Š Hyponex š¸Éž¦³„°—oª¥œÊ嘵¨¤°¨Ã˜­ ‡ªµ¤Á o¤ oœ 20 „¦´¤˜n°¨·˜¦ ¨³­µ¦‡ª‡»¤ „µ¦Á˜·Ã˜ Kn ‡ªµ¤Á o¤ oœ 4.65 ŤÇ¦Ã¤¨µ¦r ¦nª¤„´ 2,4-D ‡ªµ¤Á o¤ oœ 4.52 ŤÇ¦Ã¤¨µ¦r (­¼˜¦ 6) (˜µ¦µŠš¸É 13) ­nªœ‹ÎµœªœÄÁŒ¨¥ °Š¥°—Á¦¸É ·É¤˜oœ ¡ªnµ ­¼˜¦°µ®µ¦—´—ž¨Š Hyponex š¸É ž¦³„°—ª¥œo Ê嘵¨Ž¼Ã‡¦­ ‡ªµ¤Á o¤ oœ 20 „¦´¤˜n°¨·˜¦ ¨³­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ Kn ‡ªµ¤ 56

Á o¤ oœ 4.65 ŤÇ¦Ã¤¨µ¦r ¦nª¤„´ 2,4-D ‡ªµ¤Á o¤ oœ 4.52 ŤÇ¦Ã¤¨µ¦r (­¼˜¦ 2) Ä®o‹ÎµœªœÄ ÁŒ¨¸É¥¤µ„š¸É­»—Ášnµ„´ 6.7 ĝ (˜µ¦µŠš¸É 13, ¦¼žš¸É 11C) ­nªœ­¼˜¦°µ®µ¦°ºÉœÇ Ä®o‹ÎµœªœÄÁŒ¨¸É¥ ¦³®ªnµŠ 5.8-6.5 ĝ

57

˜µ¦µŠš¸É 12 Ÿ¨ °Š­¼˜¦°µ®µ¦—´—ž¨Š Hyponex ¦nª¤„œ´ Ê嘵¨ 3 œ·— ¨³­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ TDZ, Kn ¨³ 2,4-D ˜n°„µ¦´„œÎµ„µ¦Á„—¥°—šª· ¸‡¼–‹µ„ ­nªœ¥°— °Š„¨oª¥Å¤ o Paphiopedilum ‘Delrosi’ ®¨´Š‹µ„„µ¦Á¡µ³Á¨¥ŠÁž¸Ê }œ¦³¥³Áª¨µ 16 ­´ž—µ® r ­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ ­¼˜¦ œÊ嘵¨ „µ¦Á‹¦·Á˜·Ã˜ °Š¥°—Ä®¤n1/ (ôM) Hyponex (20 g/l) TDZ Kn 2,4-D „µ¦Á„·—¥°—3/ (%) ‹Îµœªœ¥°—2/ œÊ宜´„­—¦ª¤2/,4/ (mg) ‹ÎµœªœÄ2/ ‹Îµœªœ¦µ„2/ 1 sucrose 0.45 4.52 80.00±13.33 a 1.50±0.48 ab 120.50±37.87 bc 2.7±0.2 a 1.1±0.4 abc

2 sucrose 4.65 4.52 70.00±15.28 a 1.60±0.43 ab 252.43±80.30 a 2.8±0.3 a 1.4±0.3 a 3 glucose 0.45 4.52 80.00±13.33 a 1.80±0.49 a 162.63±61.93 ab 2.4±0.3 a 1.3±0.3 ab 4 glucose 4.65 4.52 70.00±15.28 a 1.40±0.50 ab 111.29±38.76 bc 2.7±0.3 a 1.2±0.3 abc 5 maltose 0.45 4.52 60.00±16.33 a 1.10±0.48 ab 28.86±16.31 c 2.2±0.3 a 0.4±0.2 c 6 maltose 4.65 4.52 50.00±16.67 a 0.50±0.17 b 23.80±8.82 c 2.8±0.6 a 0.4±0.2 bc 1/˜´ªÁ¨ (‡nµÁŒ¨¸É¥ ± S.E.) š¸É˜µ¤—oª¥˜ ´ª°´„¬¦Á®¤º°œ„´œÄœÂœª—·ÉŠ ­—ŠªnµÅ¤n¤¸‡ªµ¤Â˜„˜nµŠšµŠ­™·˜·š¸É¦³—´‡ªµ¤ÁºÉ°¤´Éœ 90 Áž°¦rÁŽÈœ˜r (P = 0.1) šÎµ„µ¦š—¨°Š 10 ŽÊε; 2/×¥ª·›¸„µ¦Áž¦¸¥Áš¸¥Â DMRT;

3/ 4/ ×¥ª·›¸„µ¦Áž¦¸¥Áš¸¥Â Kruskal-Wallis H ¨³ Mann-Whitney U; œÊ宜´„­—¦ª¤ = œÊ宜´„­—˜oœ¦ª¤„´œÊ宜´„­—¦µ„

58

˜µ¦µŠš¸É 13 Ÿ¨ °Š­¼˜¦°µ®µ¦—´—ž¨Š Hyponex ¦nª¤„œ´ Ê嘵¨ 3 œ·— ¨³­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ TDZ, Kn ¨³ 2,4-D ˜n°„µ¦Á‹¦·Á˜·Ã˜ °Š¥°—Á¦·É¤˜oœ °Š„¨oª¥Å¤ o Paphiopedilum ‘Delrosi’ ®¨´Š‹µ„„µ¦Á¡µ³Á¨¸Ê¥ŠÁžœ¦³¥³Áª¨µ} 16 ­´ž—µ® r ­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ ­¼˜¦ œÊ嘵¨ „µ¦Á‹¦·Á˜·Ã˜ °Š¥°—Á¦·É¤˜oœ1/ (ôM) Hyponex (20 g/l) TDZ Kn 2,4-D œÊ宜´„­—¦ª¤2/,4/ (mg) œÊ宜´„­—¥°—2/ (mg) œÊ宜´„­—¦µ„2/ (mg) ‹ÎµœªœÄ2/ ‹Îµœªœ¦µ„2/ 1 sucrose 0.45 4.52 1830.00±117.97 a 1139.80±87.96 a 690.20±58.74 a 6.5±0.3 ab 5.9±0.3 a

2 sucrose 4.65 4.52 1974.60±112.84 a 1129.20±86.48 a 845.40±65.31 a 6.7±0.2 a 5.4±0.3 a 3 glucose 0.45 4.52 1812.00±203.98 a 1068.40±123.68 a 743.60±96.17 a 6.5±0.2 ab 5.9±0.4 a 4 glucose 4.65 4.52 1995.20±83.09 a 1163.50±59.27 a 831.70±58.22 a 6.4±0.2 ab 5.6±0.3 a 5 maltose 0.45 4.52 914.60±117.19 b 725.30±96.42 b 189.30±28.79 b 6.2±0.3 ab 3.6±0.5 b 6 maltose 4.65 4.52 1137.10±75.87 b 945.30±55.92 a 191.80±25.19 b 5.8±0.5 b 2.5±0.3 c 1/˜´ªÁ¨ (‡nµÁŒ¨¸É¥ ± S.E.) š¸É˜µ¤—oª¥˜ ´ª°´„¬¦Á®¤º°œ„´œÄœÂœª—·ÉŠ ­—ŠªnµÅ¤n¤¸‡ªµ¤Â˜„˜nµŠšµŠ­™·˜·š¸É¦³—´‡ªµ¤ÁºÉ°¤´Éœ 90 Áž°¦rÁŽÈœ˜r (P = 0.1) šÎµ„µ¦š—¨°Š 10 ŽÊε; 2/×¥ª·›¸„µ¦Áž¦¸¥Áš¸¥Â DMRT;

3/ 4/ ×¥ª·›¸„µ¦Áž¦¸¥Áš¸¥Â Kruskal-Wallis H ¨³ Mann-Whitney U; œÊ宜´„­—¦ª¤ = œÊ宜´„­—˜oœ¦ª¤„´œÊ宜´„­—¦µ„

59

£µ¡š¸É 11 Ÿ¨ °Š­¼˜¦°µ®µ¦—´—ž¨Š Hyponex ¦nª¤„´œÊ嘵¨ 3 œ·— ¨³­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ TDZ, Kn ¨³ 2,4-D ˜n°„µ¦Á‹¦·Á˜·Ã˜Â¨³„µ¦´„œÎµ„µ¦Á„·—¥°—šª¸‡¼–‹µ„­nªœ¥°— °Š„¨oª¥Å¤o Paphiopedilum ‘Delrosi’ ®¨´Š‹µ„„µ¦Á¡µ³Á¨¸Ê¥ŠÁž}œ¦³¥³Áª¨µ 16 ­´ž—µ®r A = ­¼˜¦°µ®µ¦—´—ž¨Š Hyponex + Ž¼Ã‡¦­ + TDZ 0.45 ôM + 2,4-D 4.52 ôM (­¼˜¦ 1) B = ­¼˜¦°µ®µ¦—´—ž¨Š Hyponex + „¨¼Ã‡­ + TDZ 0.45 ôM + 2,4-D 4.52 ôM (­¼˜¦ 3) C = ­¼˜¦°µ®µ¦—´—ž¨Š Hyponex + Ž¼Ã‡¦­ + Kn 4.65 ôM + 2,4-D 4.52 ôM (­¼˜¦ 2) 60

3. «¹„¬µ­¼˜¦°µ®µ¦š¸ÉÁ®¤µ³­¤˜n°„µ¦´„œÎµ„µ¦Á„·—¥°—šª¸‡¼–‹µ„˜œ„¨o oª¥Å¤o Paphiopedilum ‘Delrosi’ š¸É˜´—­nªœ¥°—°°„ œÎµ˜oœ„¨ª¥Å¤o o Paphiopedilum ‘Delrosi’ Á¡µ³Á¨¸Ê¥ŠÄœš¸É¤º—Áž}œÁª¨µ 16 ­´ž—µ®r Á¡ºÉ°Ä®o o° ÁœºÊ°Á¥ºÉ°¦³®ªnµŠ o° ¨³Äš¸É¦·Áª–¥°—¤¸‡ªµ¤¥µª¡°š¸É‹³˜´—°°„Å—o ‹µ„œ´Êœ œÎµ˜oœš¸É˜´—­nªœ ¥°—°°„ ŞÁ¡µ³Á¨¸Ê¥Šœ°µ®µ¦­´ŠÁ‡¦µ³®r—´—ž¨Š­¼˜¦ Hyponex [ž»l¥ Hyponex (6.5N-4.5P-19K) ‡ªµ¤Á o¤ oœ 3 „¦´¤˜n°¨·˜¦ + ÁžžÃ˜œ (peptone) ‡ªµ¤Á o¤ oœ 2 „¦´¤˜n°¨·˜¦ + œÊε˜o¤¤´œ ¦´ÉŠ 100 „¦´¤˜n°¨·˜¦] š¸Éž¦³„°—ª¥œo Ê嘵¨ 3 œ·— ‡º° Ž¼Ã‡¦­ „¨¼Ã‡­ ¨³¤°¨Ã˜­ ‡ªµ¤Á o¤ oœ 20 „¦´¤ ˜n°¨·˜¦ ¦nª¤„´­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ TDZ ‡ªµ¤Á o¤ oœ 0.45 ŤÇ¦Ã¤¨µ¦r ®¦º° TDZ ‡ªµ¤ Á o¤ oœ 0.45 ŤÇ¦Ã¤¨µ¦r ¦nª¤„´ 2,4-D ‡ªµ¤Á o¤ oœ 4.52 ŤÇ¦Ã¤¨µ¦r ®¦º° Kn ‡ªµ¤Á o¤ oœ 4.65 ŤÇ¦Ã¤¨µ¦r ¦nª¤„´ 2,4-D ‡ªµ¤Á ¤ o oœ 4.52 ŤÇ¦Ã¤¨µ¦r —´Š˜µ¦µŠš¸É 4 Áž}œ¦³¥³Áª¨µ 16 ­´ž—µ®r ¡ªnµ š»„­¼˜¦°µ®µ¦Ä®oÁž°¦rÁŽÈœ˜r„µ¦¦°—¸ª·˜ °Š¥°—Á¦·É¤˜oœ Áž°¦rÁŽÈœ˜r„µ¦Á„·—¥°—

Ä®¤n ‹Îµœªœ¥°—Ä®¤n ¨³‹ÎµœªœÄ¥°—Ä®¤nÁŒ¨¥Å¤¸É ˜„˜n nµŠ„´œ°¥µŠ¤n ¸œ´¥­Îµ‡´šµŠ­™·˜·š¸ÉnªŠ ‡ªµ¤ÁºÉ°¤´Éœ 90 Áž°¦rÁŽÈœ˜r °¥nµŠÅ¦„Șµ¤ ­¼˜¦°µ®µ¦—´—ž¨Š Hyponex š¸Éž¦³„°—ª¥œo Ê嘵¨

¤°¨Ã˜­ ‡ªµ¤Á o¤ oœ 20 „¦´¤˜n°¨·˜¦ š¸ÉŤn¤¸­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ (­¼˜¦ 9) ®¦º°¦nª¤„´­µ¦ ‡ª‡»¤„µ¦Á˜Ã˜· TDZ ‡ªµ¤Á o¤ oœ 0.45 ŤÇ¦Ã¤¨µ¦r (­¼˜¦ 10) Ä®oÁž°¦rÁŽÈœ˜r„µ¦Á„·—¥°—ÁŒ¨¸É¥

100 Áž°¦rÁŽÈœ˜r Ä®o‹Îµœªœ¥°—Ä®¤nÁŒ¨¸É¥ 1.5 ¨³ 1.6 ¥°— ˜µ¤¨Îµ—´ (˜µ¦µŠš ¸É 14, ¦¼žš¸É 12C ¨³

12D) ˜n¡ªnµ­¼˜¦°µ®µ¦—´—ž¨Š Hyponex š¸Éž¦³„° —oª¥œÊ嘵¨¤°¨Ã˜­š»„­¼˜¦°µ®µ¦Ä®o ¥°—Ä®¤n¤¸œÊ宜´„­—¦ª¤ ¨³‹Îµœªœ¦µ„ÁŒ¨ ¸É¥œo°¥ ¨³¦µ„¤¸¨´„¬–³­´ÊœŸ·—ž„˜ · ¤¸ œ¦µ„œo°¥ ®¦º°Å¤n¤¸ œ¦µ„ Ĝ –³š¸É­¼˜¦°µ®µ¦—´—ž¨Š Hyponex š¸Éž¦³„°—oª¥œÊ嘵¨Ž¼Ã‡¦­ ‡ªµ¤ Á o¤ oœ 20 „¦´¤˜n°¨·˜¦ ¨³­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ Kn ‡ªµ¤Á o¤ oœ 4.65 ŤÇ¦Ã¤¨µ¦r ¦nª¤„´ 2,4-D ‡ªµ¤Á o¤ oœ 4.52 ŤÇ¦Ã¤¨µ¦r (­¼˜¦ 4) Ä®oÁž°¦ÁŽr Ȝ˜r„µ¦Á„—¥°—Ä®¤· nÁŒ¨¸É¥ 90 Áž°¦rÁŽÈœ˜ r ¨³Ä®o‹Îµœªœ¥°—Ä®¤nÁŒ¨¸É¥ 1.6 ¥°— Á¤ºÉ°Áž¦¸¥Áš¸¥„´­¼˜¦°µ®µ¦—´—ž¨Š Hyponex š¸É ž¦³„°—ª¥œo Ê嘵¨Ž¼Ã‡¦­Â¨³„¨¼Ã‡­ (˜µ¦µŠš¸É 14, ¦¼žš¸É 12B) ­nªœœÊ宜´„­—¦ª¤ÁŒ¨¸É¥ ¡ªnµ ­¼˜¦°µ®µ¦—´—ž¨Š Hyponex š¸Éž¦³„°—oª¥œÊ嘵¨Ž¼Ã‡¦­ ‡ªµ¤Á o¤ oœ 20 „¦´¤˜n°¨·˜¦ ¦nª¤„´ ­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ TDZ ‡ªµ¤Á o¤ oœ 0.45 ŤÇ¦Ã¤¨µ¦r ¦nª¤„´ 2,4-D ‡ªµ¤Á o¤ oœ 4.52 Ť Ǧ䨵¦r (­¼˜¦ 3) Ä®oœÊ宜´„­— °Š¥°—Ä®¤n¤¸œÊ宜„ÁŒ¨´ ¸É¥¤µ„š¸É­»— 606.86 ¤·¨¨·„¦´¤ (˜µ¦µŠš¸É 14, ¦¼žš¸É 12A) ¨³š»„­¼˜¦°µ®µ¦Ä®o‹ÎµœªœÄ¥°—Ä®¤nÁŒ¨¸É¥Å¤Â˜„˜n nµŠ„´œ°¥nµŠ¤¸œ´¥­Îµ‡´šµŠ ­™·˜·š¸ÉnªŠ‡ªµ¤ÁºÉ°¤´Éœ 90 Áž°¦rÁŽÈœ˜r

61

˜µ¦µŠš¸É 14 Ÿ¨ °Š­¼˜¦°µ®µ¦—´—ž¨Š Hyponex ¦nª¤„œ´ Ê嘵¨ 3 œ·— ¨³­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ TDZ, Kn ¨³ 2,4-D ˜n°„µ¦´„œÎµ„µ¦Á„—¥°—šª· ¸‡¼–‹µ„ ˜oœ„¨oª¥Å¤ o Paphiopedilum ‘Delrosi’ š¸É˜´—­nªœ¥°—°°„ ®¨´Š‹µ„„µ¦Á¡µ³Á¨¸Ê¥ŠÁžœ¦³¥³Áª¨µ} 16 ­´ž—µ® r ­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ ­¼˜¦ œÊ嘵¨ „µ¦¦°—¸ª·˜˜oœ „µ¦Á‹¦·Á˜·Ã˜ °Š¥°—Ä®¤n1/ (ôM) Hyponex (20 g/l) Á—·¤2/ (%) TDZ Kn 2,4-D „µ¦Á„·—¥°—3/(%) ‹Îµœªœ¥°—3/ œÊ宜´„­—¦ª¤2/,4/(mg) ‹ÎµœªœÄ3/ ‹Îµœªœ¦µ„2/ 1 sucrose 90.00±10.00 a 60.00±16.33 a 1.20±0.42 a 383.78±163.60 bc 3.7±0.3 a 2.8±0.7 abcd

2 sucrose 0.45 90.00±10.00 a 80.00±13.33 a 1.10±0.28 a 392.75±126.60 bc 3.8±0.3 a 3.1±0.7 abc 3 sucrose 0.45 4.52 90.00±10.00 a 70.00±15.28 a 1.60±0.52 a 606.86±132.01 a 4.1±0.3 a 3.3±0.4 ab 4 sucrose 4.65 4.52 90.00±10.00 a 90.00±10.00 a 1.60±0.45 a 347.78±95.41 c 4.0±0.1 a 3.3±0.5 ab 5 glucose 90.00±10.00 a 80.00±13.33 a 0.90±0.18 a 564.25±109.11 ab 4.1±0.1 a 3.9±0.2 a 6 glucose 0.45 100.00±0.00 a 70.00±15.28 a 1.20±0.36 a 288.29±62.51 cd 3.8±0.2 a 3.1±0.2 abc

7 glucose 0.45 4.52 100.00±0.00 a 70.00±15.28 a 1.50±0.58 a 134.00±38.67 d 3.5±0.2 a 2.2±0.4 cde 8 glucose 4.65 4.52 90.00±10.00 a 80.00±13.33 a 1.00±0.21 a 273.75±59.85 cd 3.6±0.2 a 2.8±0.4 bcd 9 maltose 80.00±13.33 a 100.00±0.00 a 1.50±0.22 a 119.70±23.97 d 3.7±0.1 a 1.9±0.2 de

10 maltose 0.45 90.00±10.00 a 100.00±0.00 a 1.60±0.34 a 84.60±14.85 d 3.6±0.3 a 1.6±0.3 e 11 maltose 0.45 4.52 80.00±13.33 a 90.00±10.00 a 0.90±0.10 a 135.11±29.72 d 3.7±0.2 a 2.0±0.3 de 12 maltose 4.65 4.52 90.00±10.00 a 80.00±13.33 a 1.00±0.26 a 89.63±18.63 d 3.7±0.2 a 1.3±0.2 e 1/˜´ªÁ¨ (‡nµÁŒ¨¸É¥ ± S.E.) š¸É˜µ¤—oª¥˜´ª°´„¬¦Á®¤º°œ„´œÄœÂœª—·ÉŠ ­—ŠªnµÅ¤n¤¸‡ªµ¤Â˜„˜nµŠšµŠ­™·˜·š¸É¦³—´‡ªµ¤ÁºÉ°¤´Éœ 90 Áž°¦rÁŽÈœ˜r (P = 0.1) šÎµ„µ¦š—¨°Š 10 ŽÊε; 2/×¥ª·›¸„µ¦Áž¦¸¥Áš¸¥Â DMRT; 3/×¥ª·›¸„µ¦Áž¦¸¥Áš¸¥Â Kruskal-Wallis H ¨³ Mann-Whitney U; 4/œÊ宜´„­—¦ª¤ = œÊ宜´„­—˜oœ¦ª¤„´œÊ宜´„­—¦µ„

62

£µ¡š¸É 12 Ÿ¨ °Š­¼˜¦°µ®µ¦—´—ž¨Š Hyponex ¦nª¤„´œÊ嘵¨ 3 œ·— ¨³­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ TDZ, Kn ¨³ 2,4-D ˜n°„µ¦´„œÎµ„µ¦Á„·—¥°—šª¸‡¼–‹µ„˜oœ„¨oª¥Å¤o Paphiopedilum ‘Delrosi’ š¸É˜´—­nªœ¥°—°°„ ®¨´Š‹µ„„µ¦Á¡µ³Á¨¸Ê¥ŠÁž}œ¦³¥³Áª¨µ 16 ­´ž—µ®r A = ­¼˜¦°µ®µ¦—´—ž¨Š Hyponex + Ž¼Ã‡¦­ + TDZ 0.45 ôM + 2,4-D 4.52 ôM (­¼˜¦ 3) B = ­¼˜¦°µ®µ¦—´—ž¨Š Hyponex + Ž¼Ã‡¦­ + Kn 4.65 ôM + 2,4-D 4.52 ôM (­¼˜¦ 4) C = ­¼˜¦°µ®µ¦—´—ž¨Š Hyponex + ¤°¨Ã˜­ (­¼˜¦ 9-12)

63

£µ¡š¸É 12 (˜n°) Ÿ¨ °Š­¼˜¦°µ®µ¦—´—ž¨Š Hyponex ¦nª¤„´œÊ嘵¨ 3 œ·— ¨³­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ TDZ, Kn ¨³ 2,4-D ˜n°„µ¦´„œÎµ„µ¦Á„·—¥°—šª¸‡¼–‹µ„˜oœ„¨oª¥Å¤o Paphiopedilum ‘Delrosi’ š¸É˜´—­nªœ¥°—°°„ ®¨´Š‹µ„„µ¦Á¡µ³Á¨¸Ê¥ŠÁž}œ¦³¥³Áª¨µ 16 ­´ž—µ®r D = ­¼˜¦°µ®µ¦—´—ž¨Š Hyponex + ¤°¨Ã˜­ + TDZ 0.45 ôM (­¼˜¦ 10)

64

4. «¹„¬µ­¼˜¦°µ®µ¦š¸ÉÁ®¤µ³­¤˜n°„µ¦´„œÎµ„µ¦Á„·—¥°—‹µ„­nªœ o° °Š„¨oª¥Å¤o Paphiopedilum ‘Delrosi’ œÎµ­nªœ o° °Š˜oœ„¨oª¥Å¤o Paphiopedilum ‘Delrosi’ ‹µ„„µ¦š—¨°Šš¸É 3 ŞÁ¡µ³Á¨¸Ê¥Š œ°µ®µ¦­´ŠÁ‡¦µ³®r—´—ž¨Š­¼˜¦ Hyponex [ž»l¥ Hyponex (6.5N-4.5P-19K) ‡ªµ¤Á o¤ oœ 3 „¦´¤ ˜n°¨·˜¦ + ÁžžÃ˜œ (peptone) ‡ªµ¤Á ¤ o oœ 2 „¦´¤˜n°¨·˜¦ + œÊε˜o¤¤´œ ¦´ÉŠ 100 „¦´¤˜n°¨·˜¦] š¸É ž¦³„°—ª¥œo Ê嘵¨ 3 œ·— ‡º° Ž¼Ã‡¦­ „¨¼Ã‡­ ¨³¤°¨Ã˜­ ‡ªµ¤Á o¤ oœ 20 „¦´¤˜n°¨·˜¦ ¦nª¤„´ ­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ TDZ ‡ªµ¤Á o¤ oœ 0.45 ŤÇ¦Ã¤¨µ¦r ®¦º° TDZ ‡ªµ¤Á o¤ oœ 0.45 ŤÇ¦ 䨵¦r ¦nª¤„´ 2,4-D ‡ªµ¤Á o¤ oœ 4.52 ŤÇ¦Ã¤¨µ¦r ®¦º° Kn ‡ªµ¤Á o¤ oœ 4.65 ŤÇ¦Ã¤¨µ¦r ¦nª¤„´ 2,4-D ‡ªµ¤Á o¤ oœ 4.52 ŤÇ¦Ã¤¨µ¦r —´Š˜µ¦µŠš¸É 5 Áž}œ¦³¥³Áª¨µ 16 ­´ž—µ®r ¡ªnµ ­¼˜¦ °µ®µ¦—´—ž¨Š Hyponex š¸Éž¦³„°—ª¥œo Ê嘵¨Ž¼Ã‡¦­ ‡ªµ¤Á o¤ oœ 20 „¦´¤˜n°¨·˜¦ š¸ÉŤn¤¸­µ¦ ‡ª‡»¤„µ¦Á˜Ã˜· (­¼˜¦ 1) ®¦º°­¼˜¦°µ®µ¦—´—ž¨Š Hyponex š¸Éž¦³„°—ª¥œo Ê嘵¨¤°¨Ã˜­

‡ªµ¤Á o¤ oœ 20 „¦´¤˜n°¨·˜¦ š¸ÉŤn¤¸­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ (­¼˜¦ 9) ®¦º°¦nª¤„´­µ¦‡ª‡»¤„µ¦ Á˜·Ã˜ TDZ ‡ªµ¤Á o¤ oœ 0.45 ŤÇ¦Ã¤¨µ¦r (­¼˜¦ 10) ®¦º°­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ TDZ ‡ªµ¤

Á o¤ oœ 0.45 ŤÇ¦Ã¤¨µ¦r ¦nª¤„´ 2,4-D ‡ªµ¤Á o¤ oœ 4.52 ŤÇ¦Ã¤¨µ¦r (­¼˜¦ 12) Ä®Áž°¦o rÁŽÈœ˜r „µ¦Á„·—¥°—ÁŒ¨¸É¥ 100 Áž°¦rÁŽÈœ˜r ¨³Ä®o‹Îµœªœ¥°—Ä®¤nÁŒ¨¸É¥ 1.0 ¥°— (˜µ¦µŠš¸É 15, ¦¼žš¸É 13A,

13E, 13F ¨³ 13G) °¥nµŠÅ¦„Șµ¤ ­¼˜¦°µ®µ¦ oµŠ˜oœÄ®oÁž°¦rÁŽÈœ˜„µ¦Á„r ·—¥°—Ä®¤n ¨³‹Îµœªœ

¥°—Ä®¤nÁŒ¨¸É¥Å¤n˜„˜nµŠ„´œ°¥nµŠ¤¸œ´¥­Îµ‡š´ ¸ÉnªŠ‡ªµ¤ÁºÉ°¤´Éœ 90 Áž°¦rÁŽÈœ˜r „´­¼˜¦°µ®µ¦­nªœ Ä®n ¥„Áªœ­o ¼˜¦°µ®µ¦—´—ž¨Š Hyponex š¸Éž¦³„°—ª¥œo µ˜µ¨„¨ÊÎ ¼Ã‡­ ‡ªµ¤Á o¤ oœ 20 „¦´¤˜n° ¨·˜¦ š¸ÉŤn¤¸­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ (­¼˜¦ 5) ®¦º°¦nª¤„­µ¦‡ª‡´ »¤„µ¦Á˜·Ã˜ TDZ ‡ªµ¤Á o¤ oœ 0.45 ŤÇ¦Ã¤¨µ¦r (­¼˜¦ 6) š¸ÉÄ®oÁž°¦rÁŽÈœ˜r„µ¦Á„—¥°—ÁŒ¨· ¸É¥¦³®ªnµŠ 40-50 Áž°¦rÁŽÈœ˜r ˜µ¤¨Îµ—´ ¨³¤¸‹Îµœªœ¥°—Ä®¤nÁŒ¨¸É¥ 0.5 ¥°— (˜µ¦µŠš¸É 15) ­Îµ®¦´œÊ宜„­—¥°—Ä®¤´ nÁŒ¨¸É¥ ¡ªnµ ­¼˜¦°µ®µ¦—´—ž¨Š Hyponex š¸Éž¦³„°—ª¥o œÊ嘵¨Ž¼Ã‡¦­ ‡ªµ¤Á o¤ oœ 20 „¦´¤˜n°¨·˜¦ ¨³­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ Kn ‡ªµ¤Á o¤ oœ 4.65 Ť Ǧ䨵¦r ¦nª¤„´ 2,4-D ‡ªµ¤Á o¤ oœ 4.52 ŤÇ¦Ã¤¨µ¦r (­¼˜¦ 4) ®¦º°­¼˜¦°µ®µ¦—´—ž¨Š Hyponex š¸Éž¦³„°—ª¥œo Ê嘵¨„¨¼Ã‡­ ‡ªµ¤Á o¤ oœ 20 „¦´¤˜n°¨·˜¦ š¸ÉŤn¤¸­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ (­¼˜¦ 5) ®¦º°¦nª¤„´­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ TDZ ‡ªµ¤Á o¤ oœ 0.45 ŤÇ¦Ã¤¨µ¦r ¦nª¤„´ 2,4-D ‡ªµ¤Á o¤ oœ 4.52 ŤÇ¦Ã¤¨µ¦r (­¼˜¦ 7) ¤¸œÊ宜´„­—ÁŒ¨¸É¥¤µ„š¸É­»—¦³®ªnµŠ 419.22-451.78 ¤·¨¨·„¦´¤ (˜µ¦µŠš¸É 15, ¦¼žš¸É 13B, 13C ¨³ 13D) ¨³­¼˜¦°µ®µ¦ oµŠ˜oœ¤¸œÊ宜´„­—¥°—Ä®¤nÁŒ¨¸É¥ ˜„˜nµŠ„´­¼˜¦°µ®µ¦—´—ž¨Š Hyponex š¸Éž¦³„°—oª¥œÊ嘵¨Ž¼Ã‡¦­ ‡ªµ¤Á o¤ oœ 20 „¦´¤˜n° ¨·˜¦ ¦nª¤„´­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ TDZ ‡ªµ¤Á o¤ oœ 0.45 ŤÇ¦Ã¤¨µ¦r (­¼˜¦ 2) ®¦º°­¼˜¦°µ®µ¦ 65

—´—ž¨Š Hyponex š¸Éž¦³„°—oª¥œÊ嘵¨¤°¨Ã˜­ ‡ªµ¤Á o¤ oœ 20 „¦´¤˜n°¨·˜¦ š¸Éž¦³„°—ª¥o ­µ¦‡ª‡»¤„µ¦Á˜·Ã˜š»„ (­¼˜¦ 9-12) °¥nµŠ¤¸œ´¥­Îµ‡´šµŠ­™·˜·š¸ÉnªŠ‡ªµ¤ÁºÉ°¤´Éœ 90 Áž°¦rÁŽÈœ˜r (˜µ¦µŠš¸É 15) ­Îµ®¦´‹ÎµœªœÄ¥°—Ä®¤nÁŒ¨¸É¥ ¡ªnµ š»„­¼˜¦°µ®µ¦¤¸‡nµÁŒ¨¸É¥Å¤n˜„˜nµŠ„´œ°¥nµŠ¤¸ œ´¥­Îµ‡´šµŠ­™·˜·š¸ÉnªŠ‡ªµ¤ÁºÉ°¤´Éœ 90 Áž°¦rÁŽÈœ˜r ­nªœ‹Îµœªœ¦µ„¥°—Ä®¤nÁŒ¨¸É¥ ¡ªnµ ­¼˜¦ °µ®µ¦—´—ž¨Š Hyponex š¸Éž¦³„°—ª¥œo Ê嘵¨Ž¼Ã‡¦­ ‡ªµ¤Á o¤ oœ 20 „¦´¤˜°¨n ·˜¦ ¨³­µ¦ ‡ª‡»¤„µ¦Á˜Ã˜· Kn ‡ªµ¤Á o¤ oœ 4.65 ŤÇ¦Ã¤¨µ¦r ¦nª¤„´ 2,4-D ‡ªµ¤Á o¤ oœ 4.52 ŤÇ¦Ã¤ ¨µ¦r (­¼˜¦ 4) Ä®o¥°—Ä®¤n¤¸‹Îµœªœ¦µ„ÁŒ¨¸É¥¤µ„š¸É­»— 3.7 ¦µ„ °¥nµŠÅ¦„Șµ¤ ¡ªnµ ­¼˜¦°µ®µ¦ oµŠ˜oœ ­nŠŸ¨Ä®o¥°—Ä®¤n¤¸‹Îµœªœ¦µ„ÁŒ¨¸É¥Å¤n˜„˜nµŠ„´­¼˜¦°µ®µ¦­nªœÄ®n°¥nµŠ¤¸œ´¥­Îµ‡´šµŠ ­™·˜·š¸ÉnªŠ‡ªµ¤ÁºÉ°¤´Éœ 90 Áž°¦rÁŽÈœ˜r ¥„Áªoœ ­¼˜¦°µ®µ¦—´—ž¨Š Hyponex š¸Éž¦³„°—oª¥œÊ嘵¨ ¤°¨Ã˜­ ‡ªµ¤Á o¤ oœ 20 „¦´¤˜n°¨·˜¦ š¸ÉŤn¤¸­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ (­¼˜¦ 9) ®¦º°¦nª¤„´­µ¦ ‡ª‡»¤„µ¦Á˜Ã˜· Kn ‡ªµ¤Á o¤ oœ 4.65 ŤÇ¦Ã¤¨µ¦r ¦nª¤„´ 2,4-D ‡ªµ¤Á o¤ oœ 4.52 ŤÇ¦Ã¤

¨µ¦r š¸ÉÄ®o‹Îµœªœ¦µ„œo°¥š¸É­»— ‡º° 2.7 ¦µ„ (­¼˜¦ 12) (˜µ¦µŠš¸É 15)

66

˜µ¦µŠš¸É 15 Ÿ¨ °Š­¼˜¦°µ®µ¦—´—ž¨Š Hyponex ¦nª¤„œ´ Ê嘵¨ 3 œ·— ¨³­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ TDZ, Kn ¨³ 2,4-D ˜n°„µ¦´„œÎµ„µ¦Á„—¥°—‹µ„­· nªœ o° °Š„¨oª¥Å¤ o Paphiopedilum ‘Delrosi’ ®¨´Š‹µ„„µ¦Á¡µ³Á¨¸Ê¥ŠÁžœ¦³¥³Áª¨µ} 16 ­´ž—µ® r ­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ ­¼˜¦ œÊ嘵¨ „µ¦¦°—¸ª·˜ °Š „µ¦Á‹¦·Á˜·Ã˜ °Š¥°—Ä®¤n1/ (ôM) Hyponex (20 g/l) ·Êœ­nªœ o°2/ (%) TDZ Kn 2,4-D „µ¦Á„·—¥°—3/ (%) ‹Îµœªœ¥°—3/ œÊ宜´„­—¦ª¤2/,4/ (mg) ‹ÎµœªœÄ3/ ‹Îµœªœ¦µ„2/ 1 sucrose 90.00±10.00 a 100.00±0.00 a 1.00±0.00 a 374.80±61.61 ab 3.7±0.2 a 3.1±0.3 ab

2 sucrose 0.45 70.00±15.28 b 80.00±13.33 abc 0.80±0.13 ab 245.38±45.38 bcd 3.4±0.2 a 2.9±0.2 ab 3 sucrose 0.45 4.52 80.00±13.33 a 80.00±13.33 abc 0.80±0.13 ab 336.88±41.94 abc 3.3±0.2 a 3.0±0.2 ab

4 sucrose 4.65 4.52 80.00±13.33 a 90.00±10.00 ab 1.00±0.15 a 419.22±65.85 a 3.9±0.4 a 3.7±0.5 a 5 glucose 30.00±15.28 bc 40.00±16.33 c 0.50±0.22 b 450.50±129.11 a 3.5±0.3 a 3.4±0.5 ab 6 glucose 0.45 30.00±15.28 bc 50.00±16.67 bc 0.50±0.17 b 338.20±122.28 abc 3.0±0.3 a 3.0±0.5 ab

7 glucose 0.45 4.52 70.00±15.28 b 90.00±10.00 ab 0.90±0.10 ab 451.78±83.53 a 3.6±0.2 a 3.6±0.3 ab 8 glucose 4.65 4.52 20.00±13.33 c 80.00±13.33 abc 0.80±0.13 ab 378.38±65.28 ab 3.4±0.2 a 3.5±0.4 ab

9 maltose 20.00±13.33 c 100.00±0.00 a 1.00±0.00 a 160.50±17.49 d 3.2±0.1 a 2.7±0.2 b

10 maltose 0.45 10.00±10.00 c 100.00±0.00 a 1.00±0.00 a 187.10±23.35 cd 3.4±0.2 a 3.0±0.2 ab 11 maltose 0.45 4.52 10.00±10.00 c 90.00±10.00 ab 0.90±0.10 ab 190.00±17.10 cd 3.4±0.2 a 3.0±0.2 ab 12 maltose 4.65 4.52 40.00±16.33 bc 100.00±0.00 a 1.00±0.00 a 156.10±22.11 d 3.3±0.2 a 2.7±0.3 b 1/˜´ªÁ¨ (‡nµÁŒ¨¸É¥ ± S.E.) š¸É˜µ¤—oª¥˜´ª°´„¬¦Á®¤º°œ„´œÄœÂœª—·ÉŠ ­—ŠªnµÅ¤n¤¸‡ªµ¤Â˜„˜nµŠšµŠ­™·˜·š¸É¦³—´‡ªµ¤ÁºÉ°¤´Éœ 90 Áž°¦rÁŽÈœ˜r (P = 0.1) šÎµ„µ¦š—¨°Š 10 ŽÊε; 2/×¥ª·›¸„µ¦Áž¦¸¥Áš¸¥Â DMRT; 3/×¥ª·›¸„µ¦Áž¦¸¥Áš¸¥Â Kruskal-Wallis H ¨³ Mann-Whitney U; 4/œÊ宜´„­—¦ª¤ = œÊ宜´„­—˜oœ¦ª¤„´œÊ宜´„­—¦µ„

67

£µ¡š¸É 13 Ÿ¨ °Š­¼˜¦°µ®µ¦—´—ž¨Š Hyponex ¦nª¤„´œÊ嘵¨ 3 œ·— ¨³­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ TDZ, Kn ¨³ 2,4-D ˜n°„µ¦´„œÎµ„µ¦Á„·—¥°—‹µ„­nªœ o° °Š„¨oª¥Å¤o Paphiopedilum ‘Delrosi’ ®¨´Š‹µ„„µ¦ Á¡µ³Á¨¸Ê¥ŠÁž}œ¦³¥³Áª¨µ 16 ­´ž—µ®r A = ­¼˜¦°µ®µ¦—´—ž¨Š Hyponex + Ž¼Ã‡¦­ (­¼˜¦ 1-4) B = ­¼˜¦°µ®µ¦—´—ž¨Š Hyponex + Ž¼Ã‡¦­ + Kn 4.65 ôM + 2,4-D 4.52 ôM (­¼˜¦ 4) C = ­¼˜¦°µ®µ¦—´—ž¨Š Hyponex + „¨¼Ã‡­ (­¼˜¦ 5-8) 68

£µ¡š¸É 13 (˜n°) Ÿ¨ °Š­¼˜¦°µ®µ¦—´—ž¨Š Hyponex ¦nª¤„´œÊ嘵¨ 3 œ·— ¨³­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ TDZ, Kn ¨³ 2,4-D ˜n°„µ¦´„œÎµ„µ¦Á„·—¥°—‹µ„­nªœ o° °Š„¨oª¥Å¤o Paphiopedilum ‘Delrosi’ ®¨´Š‹µ„„µ¦ Á¡µ³Á¨¸Ê¥ŠÁž}œ¦³¥³Áª¨µ 16 ­´ž—µ®r D = ­¼˜¦°µ®µ¦—´—ž¨Š Hyponex + „¨¼Ã‡­ + TDZ 0.45 ôM + 2,4-D 4.52 ôM (­¼˜¦ 7) E = ­¼˜¦°µ®µ¦—´—ž¨Š Hyponex + ¤°¨Ã˜­ (­¼˜¦ 9-12) F = ­¼˜¦°µ®µ¦—´—ž¨Š Hyponex + ¤°¨Ã˜­ + TDZ 0.45 ôM (­¼˜¦ 10) 69

£µ¡š¸É 13 (˜n°) Ÿ¨ °Š­¼˜¦°µ®µ¦—´—ž¨Š Hyponex ¦nª¤„´œÊ嘵¨ 3 œ·— ¨³­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ TDZ, Kn ¨³ 2,4-D ˜n°„µ¦´„œÎµ„µ¦Á„·—¥°—‹µ„­nªœ o° °Š„¨oª¥Å¤o Paphiopedilum ‘Delrosi’ ®¨´Š‹µ„„µ¦ Á¡µ³Á¨¸Ê¥ŠÁž}œ¦³¥³Áª¨µ 16 ­´ž—µ®r

G = ­¼˜¦°µ®µ¦—´—ž¨Š Hyponex + ¤°¨Ã˜­ + TDZ 0.45 ôM + 2,4-D 4.52 ôM (­¼˜¦ 11)

70

5. «¹„¬µ­¼˜¦°µ®µ¦š¸ÉÁ®¤µ³­¤˜n°„µ¦´„œÎµ„µ¦Á„·—‡¨¨´­ ®¦º° somatic embryos ‹µ„·ÊœÁœºÊ° Á¥ºÉ°¦³®ªnµŠ o° °Š„¨oª¥Å¤ o Paphiopedilum ‘Delrosi’ œÎµ·ÊœÁœºÊ°Á¥ºÉ°¦³®ªnµŠ o° °Š„¨oª¥Å¤o Paphiopedilum ‘Delrosi’ ‹µ„„µ¦š—¨°Šš¸É 3 ¤µ ˜´—Ä®o¤¸‡ªµ¤¥µª 0.5 ÁŽœ˜·Á¤˜¦ ŞÁ¡µ³Á¨¸Ê¥Šœ­¼˜¦°µ®µ¦ 1/2 MS (1/2 strength macro-and full- strength micro elements) š¸Éž¦³„°—ª¥œo Ê嘵¨ 3 œ·— ‡º° Ž¼Ã‡¦­ „¨¼Ã‡­ ¨³¤°¨Ã˜­ ‡ªµ¤ Á o¤ oœ 20 „¦´¤˜n°¨·˜¦ ¦nª¤„´­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ TDZ ‡ªµ¤Á o¤ oœ 0.45 ¨³ 4.54 ŤÇ¦Ã¤ ¨µ¦r Á¡¸¥Š°¥nµŠÁ—¸¥ª ®¦º°¦nª¤„´ 2,4-D ‡ªµ¤Á o¤ oœ 4.52 ¨³ 13.56 ŤÇ¦Ã¤¨µ¦r —´Š˜µ¦µŠš¸É 6 Áž}œ¦³¥³Áª¨µ 16 ­´ž—µ® r ¡ªnµ ·ÊœÁœºÊ°Á¥ºÉ°¦³®ªnµŠ o°š¸ÉÁ¡µ³Á¨¸Ê¥Šœš»„­¼˜¦°µ®µ¦ oµŠ˜oœÅ¤n¤¸ „µ¦Á„·—‡¨¨´­ ®¦º° somatic embryos Á¨¥ ÁœºÉ°Š‹µ„·ÊœÁœºÊ°Á¥ºÉ°¦³®ªnµŠ o°¤¸„µ¦Áž¨¸É¥œÂž¨ŠÁžœ­} ¸ œÊ嘵¨Â¨³˜µ¥®¨´Š‹µ„Á¦·É¤Á¡µ³Á¨¸Ê¥ŠÁžœ¦} ³¥³Áª¨µž¦³¤µ– 4-5 ­´ž—µ®r (¦¼žš¸É 14A )

71

£µ¡š¸É 14 Ÿ¨ °Š­¼˜¦°µ®µ¦—´—ž¨Š Hyponex ¦nª¤„´œÊ嘵¨ 3 œ·— ¨³­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ TDZ ¨³ 2,4-D ˜n°„µ¦´„œÎµ„µ¦Á„·—‡¨¨´­ ®¦º° somatic embryos ‹µ„·ÊœÁœºÊ°Á¥ºÉ°¦³®ªnµŠ o° °Š„¨oª¥Å¤o

Paphiopedilum ‘Delrosi’ A = ·ÊœÁœºÊ°Á¥ºÉ°¦³®ªnµŠ o°š¸É¤¸„µ¦Áž¨¸É¥œÂž¨ŠÁž}œ­¸œÊ嘵¨Â¨³˜µ¥

72

6. «¹„¬µ­¼˜¦°µ®µ¦š¸ÉÁ®¤µ³­¤˜n°„µ¦´„œÎµ„µ¦Á„·—‡¨¨´­ ®¦º° somatic embryos ‹µ„·Êœ­nªœÄ °Š„¨oª¥Å¤ o Paphiopedilum ‘Delrosi’ œÎµÄ¦·Áª–¥°—‹µ„˜œ„¨o oª¥Å¤o Paphiopedilum ‘Delrosi’ ¤µ˜´—ĝ­ªœ‡¦n ¹ÉŠ¨nµŠÄ®o¤¸ œµ— 0.3 x 0.4 ˜µ¦µŠÁŽœ˜·Á¤˜¦ ŞÁ¡µ³Á¨¸Ê¥Šœ­¼˜¦°µ®µ¦ 1/2 MS (1/2 strength macro-and full-strength micro elements) š¸Éž¦³„°—oª¥œÊ嘵¨ 3 œ·— ‡º° Ž¼Ã‡¦­ „¨¼Ã‡­ ¨³¤°¨Ã˜­ ‡ªµ¤Á o¤ oœ 20 „¦´¤˜n°¨·˜¦ ¦nª¤„´­µ¦‡ª‡»¤„µ¦Á˜Ã˜· TDZ ‡ªµ¤Á o¤ oœ 0.45 ¨³ 4.54 Ť Ǧ䨵¦r Á¡¸¥Š°¥nµŠÁ—¸¥ª ®¦º°¦nª¤„´ 2,4-D ‡ªµ¤Á o¤ oœ 4.52 ¨³ 13.56 ŤÇ¦Ã¤¨µ¦r —´Š ˜µ¦µŠš¸É 7 Áž}œ¦³¥³Áª¨µ 16 ­´ž—µ®r ¡ªnµ ·Êœ­nªœÄšÁ¡µ³Á¨¸É ¸Ê¥Šœš»„­¼˜¦°µ®µ¦ oµŠ˜oœÅ¤n¤¸„µ¦ Á„·—‡¨¨´­ ®¦º° somatic embryos Á¨¥ ÁœºÉ°Š‹µ„·Êœ­ªœÄ¤n „µ¦Áž¨¸ ¸É¥œÂž¨ŠÁž}œ­¸œÊ嘵¨Â¨³˜µ¥ ®¨´Š‹µ„Á¦·É¤Á¡µ³Á¨¸Ê¥ŠÁžœ¦³¥³Áª¨µž¦³¤µ–} 1-2 ­´ž—µ® r (¦¼žš¸É 15A )

73

£µ¡š¸É 15 Ÿ¨ °Š­¼˜¦°µ®µ¦—´—ž¨Š Hyponex ¦nª¤„´œÊ嘵¨ 3 œ·— ¨³­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ TDZ ¨³ 2,4-D ˜n°„µ¦´„œÎµ„µ¦Á„·—‡¨¨´­ ®¦º° somatic embryos ‹µ„·Êœ­nªœÄ °Š„¨oª¥Å¤o Paphiopedilum

‘Delrosi’ A = ·Êœ­nªœÄš¸É¤¸„µ¦Áž¨¸É¥œÂž¨ŠÁž}œ­¸œÊ嘵¨Â¨³˜µ¥

šš¸É 5

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1. «¹„¬µ­¼˜¦°µ®µ¦š¸ÉÁ®¤µ³­¤˜n°„µ¦´„œÎµ„µ¦Á„·—¥°—šª¸‡¼–‹µ„­ªœ¥°— °Š„¨n oª¥Å¤ o Paphiopedilum ‘Delrosi’ 1.1 «¹„¬µŸ¨ °Š­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ TDZ, BA, Kn, 2,4-D ¨³ Ads ˜n°„µ¦´„œÎµ„µ¦Á„·— ¥°—šª¸‡¼–‹µ„­nªœ¥°— °Š„¨oª¥Å¤ o Paphiopedilum ‘Delrosi’ ‹µ„„µ¦«¹„¬µŸ¨ °Š­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ TDZ, BA, Kn, 2,4-D ¨³ Ads ˜n°„µ¦´„

œÎµ„µ¦Á„—¥°—šª· ¸‡¼–‹µ„­ªœ¥°— °Š„¨n oª¥Å¤ o Paphiopedilum ‘Delrosi’ ×¥ TDZ, BA, Kn ¨³ Ads Ž¹ÉŠÁž}œ­µ¦š¸É°¥¼nĜ„¨»n¤ÅŽÃ˜Å‡œ·œ (cytokinins) ¤¸šµš­Îµ‡´˜n°„µ¦´„œÎµÄ®oÁ„·—¥°— ¨³

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Hong et al. (2008b) š¸É«¹„¬µ„µ¦ ¥µ¥¡œ›´ »r„¨oª¥Å¤o Paphiopedilum ‘Alma Gavaert’ ‹µ„ PLBs ×¥ ­µ¤µ¦™´„œÎµÄ®oÁ„·—¥°—ÁŒ¨¸É¥ 3 ¥°— ‹µ„­¼˜¦°µ®µ¦š¸Éž¦³„°—ª¥­µ¦‡ª‡o »¤„µ¦Á˜·Ã˜ Kn ‡ªµ¤Á o¤ oœ 4.65 ŤÇ¦Ã¤¨µ¦r ®¨´Š‹µ„Á¡µ³Á¨¸Ê¥Š¥°—°n°œÁž}œÁª¨µ 60 ª´œ ¨³‹µ„„µ¦š—¨°Š °Š Ng and Saleh (2010a) š¸É«¹„¬µ„µ¦ ¥µ¥¡œ›´ »r„¨oª¥Å¤o Paphiopedilum rothschildianum Ž¹ÉŠ ­µ¤µ¦™´„œÎµ„µ¦Á„·— secondary PLBs ÁŒ¨¸É¥¤µ„š¸É­»— 4.1 PLBs ˜n° 1 primary PLBs ‹µ„­¼˜¦ °µ®µ¦šž¦³„°—¸É oª¥­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ Kn ‡ªµ¤Á o¤ oœ 4.0 ŤÇ¦Ã¤¨µ¦r Ĝ¦³¥³Áª¨µ 8 ­´ž—µ®r ˜˜„˜n nµŠ„´„µ¦š—¨°Š °Š Yan et al. (2006) š¸É«¹„¬µ„µ¦ ¥µ¥¡´œ›»r„¨oª¥Å¤o Cypripedium flavam ‹µ„„µ¦´„œÎµÄ®Á„o ·—¥°—šª¸‡¼–‹µ„˜oœ°n°œ ×¥­µ¤µ¦™´„œÎµÄ®oÁ„—¥°· — šª¸‡¼–¤µ„š¸É­»— 95 Áž°¦rÁŽÈœ˜r ¨³¤¸¥°—Á„—¤µ„š· ¸É­»— 2.55 ¥°— ‹µ„­¼˜¦°µ®µ¦š¸Éž¦³„°—oª¥­µ¦ ‡ª‡»¤„µ¦Á˜Ã˜· BAP ‡ªµ¤Á o¤ oœ 2.22 ŤÇ¦Ã¤¨µ¦r ¦nª¤„´œÊ嘤¤o ´œ ¦´ÉŠ ž¦·¤µ– 20 „¦´¤˜n° ¨·˜¦ ¨³„µ¦š—¨°Š °Š Chen et al. (2002) š¸É«¹„¬µ„µ¦Á„·—¥°—šª¸‡¼–‹µ„˜µ °Š„oµœn°—°„ „¨oª¥Å¤o Paphiopedilum philippinense 2 ­µ¥¡´œ›»r (PH59 ¨³ PH60) Ĝ­µ¥¡´œ›»r PH59 ¤¸·Êœ

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1.2 «¹„¬µŸ¨ °Š­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ TDZ, BA, Kn ¨³ 2,4-D ˜n°„µ¦´„œÎµ„µ¦Á„·—¥°— šª¸‡¼–‹µ„­nªœ¥°— °Š„¨oª¥Å¤ o Paphiopedilum ‘Delrosi’ „µ¦š—¨°Šœ¸ÊÁž}œ„µ¦ž¦´‡ªµ¤Á o¤ oœ °Š­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ ×¥°oµŠ°·Š„´„µ¦ š—¨°Šš¸É 1.1 Á¡ºÉ°«¹„¬µŸ¨ °Š­µ¦‡ª‡»¤„µ¦Á˜·Ã˜˜°„µ¦n ´„œÎµ„µ¦Á„·—¥°—šª¸‡¼–‹µ„­ªœ¥°—n °Š„¨oª¥Å¤ o Paphiopedilum ‘Delrosi’ Ä®o¤¸ž¦³­š›· ·£µ¡¤µ„š¸É­»— ‹µ„„µ¦Á¡·É¤®¦º°¨—‡ªµ¤ Á o¤ oœ °Š­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ TDZ, BA, Kn ¨³ 2,4-D ¡ªnµ ­¼˜¦°µ®µ¦—´—ž¨Š Hyponex š¸Éž¦³„°—ª¥­µ¦‡ª‡o »¤„µ¦Á˜·Ã˜ TDZ ‡ªµ¤Á o¤ oœ 0.45 ŤÇ¦Ã¤¨µ¦r ¦nª¤„´ 2,4-D ‡ªµ¤ Á o¤ oœ 4.52 ŤÇ¦Ã¤¨µ¦r Ä®oÁž°¦rÁŽÈœ˜r„µ¦Á„·—¥°—¨³‹Îµœªœ¥°—Ä®¤nÁŒ¨¥¤µ„¨³­¤¸É ÉεÁ­¤° š¸É­»— 90 Áž°¦rÁŽÈœ˜r ¨³¥°—Ä®¤n 1.3 ¥°— Ž¹ÉŠ‡¨oµ¥„´„µ¦š—¨°Š °Š Lin et al. (2000) š¸É«¹„¬µ„µ¦ ¥µ¥¡œ›´ »r„¨oª¥Å¤o Paphiopedilum hybrid (Paphiopedilum callosum ‘Oakhi’ x Paph. Lawreneanum ‘Tradition’) ‹µ„„µ¦Á¡µ³Á¨¸Ê¥ŠÂ‡¨¨´­ œ­¼˜¦°µ®µ¦ 1/2 MS š¸Éž¦³„°—ª¥­µ¦o 76

‡ª‡»¤„µ¦Á˜Ã˜· 2,4-D ‡ªµ¤Á o¤ oœ 5 ¤·¨¨·„¦´¤˜n°¨·˜¦ ¨³ TDZ ‡ªµ¤Á o¤ oœ 1 ¤·¨¨·„¦´¤˜n° ¨·˜¦ Ž¹ÉŠ­µ¤µ¦™¡´•œµÁž}œ PLBs ¨³˜oœš­¤¸É ¼¦–r

2. «¹„¬µœ·— °ŠœÊ嘵¨š¸ÉÁ®¤µ³­¤˜n°„µ¦´„œÎµ„µ¦Á„—¥°—šª· ¸‡¼–‹µ„­nªœ¥°— °Š„¨oª¥Å¤ o Paphiopedilum ‘Delrosi’ ‹µ„„µ¦«¹„¬µŸ¨ °ŠœÊ嘵¨ 3 œ·— ‡º° Ž¼Ã‡¦­ „¨¼Ã‡­ ¨³¤°¨Ã˜­ ‡ªµ¤Á o¤ oœ 20 „¦´¤˜n°¨·˜¦ ¦nª¤„´­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ TDZ ‡ªµ¤Á o¤ oœ 0.45 ŤÇ¦Ã¤¨µ¦r ¦nª¤„´ 2,4-D ‡ªµ¤Á o¤ oœ 4.52 ŤÇ¦Ã¤¨µ¦r ®¦º° Kn ‡ªµ¤Á o¤ oœ 4.65 ŤÇ¦Ã¤¨µ¦r ¦nª¤„´ 2,4-D ‡ªµ¤ Á o¤ oœ 4.52 ŤÇ¦Ã¤¨µ¦r ˜n°„µ¦´„œÎµ„µ¦Á„·—¥°—šª¸‡¼–‹µ„­ªœ¥°— °Š„¨n oª¥Å¤o Paphiopedilum ‘Delrosi’ ‹µ„Ÿ¨„µ¦ª·Á‡¦µ³®ršµŠ­™·˜·¸ÊÄ®oÁ®ÈœªnµÁž°¦rÁŽÈœ˜„µ¦Á„r ·—¥°—ÁŒ¨¸É¥ ‹Îµœªœ¥°—ÁŒ¨¸É¥­nªœÄ®n ¨³‹ÎµœªœÄ¥°—Ä®¤nÁŒ¨¸É¥Å¤n˜„˜nµŠ„´œ°¥nµŠ¤¸œ´¥­Îµ‡´ ˜n°¥nµŠÅ¦„È ˜µ¤ ¡ªnµ­¼˜¦°µ®µ¦š¸Éž¦³„°—ª¥œo Ê嘵¨¤°¨Ã˜­Ä®oÁž°¦rÁŽÈœ˜r„µ¦Á„·—¥°—ÁŒ¨¸É¥ ‹Îµœªœ¥°—

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¡´•œµš¸É—¸ ˜n˜nµŠ‹µ„„µ¦š—¨°Š °Š Rahman et al. (2005) š¸É«¹„¬µœÊ嘵¨ 3 œ·— (Ž¼Ã‡¦­, ¤°¨ Ø­ ‡ªµ¤Á ¤ o oœ 20 „¦´¤˜n°¨·˜¦ ¨³Ž°·š°¨ ‡ªµ¤Á o¤ oœ 10 „¦´¤˜n°¨·˜¦) ˜n°„µ¦Á¡·É¤ž¦·¤µ– ‡¨¨´­Â¨³„µ¦´„œÎµÄ®Á„o ·—˜oœš¸É­¤¼¦– °Š„¨r oª¥Å¤o Doritaenopsis ¡ªnµ ‡¨¨­š´ ¸ÉÁ¡µ³Á¨¸Ê¥Šœ ­¼˜¦°µ®µ¦ NP š¸É¤¸œÊ嘵¨¤°¨Ã˜­ ¤¸­¸Á ¸¥ªÂ¨³¤¸„µ¦Á„—Áž· }œ PLBs ¤µ„ªnµœÊ嘵¨Ž°¦r·š°¨ ˜n œÊ嘵¨Ž°¦r·š°¨ ­nŠŸ¨Ä®o PLBs ¡´•œµÁž}œ˜oœš¸É­¤¦–¼ rŗo—¸„ªnµœÊ嘵¨¤°¨Ã˜­Â¨³Ž¼Ã‡¦­ ˜n °¥nµŠÅ¦„Șµ¤ œÊ嘵¨¤°¨Ã˜­šÎµÄ®o˜oœ°n°œ¤¸„µ¦Á‹¦·Á˜·Ã˜Â¨³¡´•œµÅ—o—¸„ªnµœÊ嘵¨Ž°¦r·š°¨ ¨³Ž¼Ã‡¦­

3. «¹„¬µ­¼˜¦°µ®µ¦š¸ÉÁ®¤µ³­¤˜n°„µ¦´„œÎµ„µ¦Á„·—¥°—šª¸‡¼–‹µ„˜œ„¨o oª¥Å¤o Paphiopedilum ‘Delrosi’ š¸É˜´—­nªœ¥°—°°„ ‹µ„„µ¦«¹„¬µŸ¨ °ŠœÊ嘵¨ 3 œ·— ‡º° Ž¼Ã‡¦­ „¨¼Ã‡­ ¨³¤°¨Ã˜­ ‡ªµ¤Á o¤ oœ 20 „¦´¤˜n°¨·˜¦ ¦nª¤„´­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ TDZ ‡ªµ¤Á o¤ oœ 0.45 ŤÇ¦Ã¤¨µ¦r ®¦º° TDZ ‡ªµ¤

Á o¤ oœ 0.45 ŤÇ¦Ã¤¨µ¦r ¦nª¤„´ 2,4-D ‡ªµ¤Á o¤ oœ 4.52 ŤÇ¦Ã¤¨µ¦r ®¦º° Kn ‡ªµ¤Á o¤ oœ 4.65 ŤÇ¦Ã¤¨µ¦r ¦nª¤„´ 2,4-D ‡ªµ¤Á o¤ oœ 4.52 ŤÇ¦Ã¤¨µ¦r ˜n°„µ¦´„œÎµ„µ¦Á„·—¥°—šª¸‡¼– ‹µ„˜oœ„¨oª¥Å¤o Paphiopedilum ‘Delrosi’ š¸É˜´—­nªœ¥°—°°„ ‹µ„Ÿ¨„µ¦ª·Á‡¦µ³®ršµŠ­™·˜·¸ÊÄ®oÁ®Èœ ªnµÁž°¦rÁŽÈœ˜„µ¦Á„r ·—¥°— ‹Îµœªœ¥°— ¨³‹ÎµœªœÄ¥°—Ä®¤nÁŒ¨¸É¥Å¤n˜„˜nµŠ„´œ°¥nµŠ¤¸œ´¥­Îµ‡´

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œÊ宜„­—¦ª¤´ ¨³‹Îµœªœ¦µ„¥°—Ä®¤ ÁŒ¨n ¸É¥œo°¥ ¨³¦µ„¤¸¨´„¬–³­´ÊœŸ·—ž„˜ · ¤¸ œ¦µ„œo°¥ ®¦º°Å¤n¤¸ œ¦µ„ —´Šœœ­´Ê ¼˜¦°µ®µ¦—´—ž¨Š Hyponex š¸Éž¦³„°—ª¥œo Ê嘵¨Ž¼Ã‡¦­ ‡ªµ¤Á o¤ oœ 20 „¦´¤˜n°¨·˜¦ ¨³­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ Kn ‡ªµ¤Á o¤ oœ 4.65 ŤÇ¦Ã¤¨µ¦r ¦nª¤„´ 2,4-D ‡ªµ¤Á o¤ oœ 4.52 ŤÇ¦Ã¤¨µ¦r Ä®oÁž°¦rÁŽÈœ˜r„µ¦Á„—¥°—Ä®¤· nÁŒ¨¸É¥ 90 Áž°¦rÁŽÈœ˜r ¨³Ä®o‹Îµœªœ ¥°—Ä®¤nÁŒ¨¸É¥ 1.6 ¥°— ‹¹ŠÁž}œ­¼˜¦°µ®µ¦š¸ÉÁ®¤µ³­¤ Ž¹ÉŠ‡¨oµ¥„´„µ¦š—¨°Š °Š Hong et al. (2008b) š¸É«¹„¬µ„µ¦ ¥µ¥¡œ›´ »r„¨oª¥Å¤o Paphiopedilum ‘Alma Gavaert’ ‹µ„ PLBs ×¥­µ¤µ¦™´„ œÎµÄ®oÁ„·—¥°—ÁŒ¨¸É¥ 3 ¥°— ‹µ„­¼˜¦°µ®µ¦š¸Éž¦³„°—ª¥­µ¦‡ª‡o »¤„µ¦Á˜·Ã˜ Kn ‡ªµ¤Á o¤ oœ 4.65 ŤÇ¦Ã¤¨µ¦r ¦nª¤„´œÊ嘵¨Ž¼Ã‡¦­ ‡ªµ¤Á o¤ oœ 20 „¦´¤˜n°¨·˜¦ ®¨´Š‹µ„Á¡µ³Á¨¸Ê¥Š¥°—°n°œ Áž}œÁª¨µ 60 ª´œ ¨³‹µ„„µ¦š—¨°Š °Š Ng and Saleh (2010a) š¸É«¹„¬µ„µ¦ ¥µ¥¡´œ›»r„¨oª¥Å¤o Paphiopedilum rothschildianum Ž¹ÉŠ­µ¤µ¦™´„œÎµ„µ¦Á„·— secondary PLBs ÁŒ¨¸É¥¤µ„š¸É­»— 4.1 PLBs ˜n° 1 primary PLBs ‹µ„­¼˜¦°µ®µ¦š¸Éž¦³„°—ª¥­µ¦‡ª‡o »¤„µ¦Á˜·Ã˜ Kn ‡ªµ¤Á o¤ oœ 4.0 ŤÇ¦Ã¤¨µ¦r ¦nª¤„´œµ˜µ¨ŽÊÎ ¼Ã‡¦­ ‡ªµ¤Á o¤ oœ 30 „¦´¤˜n°¨·˜¦ Ĝ¦³¥³Áª¨µ 8 ­´ž—µ® r

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4. «¹„¬µ­¼˜¦°µ®µ¦š¸ÉÁ®¤µ³­¤˜n°„µ¦´„œÎµ„µ¦Á„·—¥°—‹µ„­nªœ o° °Š„¨oª¥Å¤o Paphiopedilum ‘Delrosi’ ‹µ„„µ¦«¹„¬µŸ¨ °ŠœÊ嘵¨ 3 œ·— ‡º° Ž¼Ã‡¦­ „¨¼Ã‡­ ¨³¤°¨Ã˜­ ‡ªµ¤Á o¤ oœ 20 „¦´¤˜n°¨·˜¦ ¦nª¤„´­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ TDZ ‡ªµ¤Á o¤ oœ 0.45 ŤÇ¦Ã¤¨µ¦r ®¦º° TDZ ‡ªµ¤ Á o¤ oœ 0.45 ŤÇ¦Ã¤¨µ¦r ¦nª¤„´ 2,4-D ‡ªµ¤Á o¤ oœ 4.52 ŤÇ¦Ã¤¨µ¦r ®¦º° Kn ‡ªµ¤Á o¤ oœ 4.65 ŤÇ¦Ã¤¨µ¦r ¦nª¤„´ 2,4-D ‡ªµ¤Á o¤ oœ 4.52 ŤÇ¦Ã¤¨µ¦r ˜n°„µ¦´„œÎµ„µ¦Á„·—¥°—‹µ„ ­nªœ o° °Š„¨oª¥Å¤ o Paphiopedilum ‘Delrosi’ ¡ªnµ ­¼˜¦°µ®µ¦—´—ž¨Š Hyponex š¸É ž¦³„°—ª¥œo Ê嘵¨Ž¼Ã‡¦­ ‡ªµ¤Á o¤ oœ 20 „¦´¤˜n°¨·˜¦ š¸ÉŤn¤¸­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ Ä®o Áž°¦rÁŽÈœ˜r„µ¦Á„·—¥°—ÁŒ¨¸É¥¤µ„š¸É­»— 100 Áž°¦rÁŽÈœ˜r ‹Îµœªœ¥°—Ä®¤nÁŒ¨¸É¥ 1.0 ¥°— ¨³œÊ宜´„ ­—¦ª¤ÁŒ¨¸É¥¥°—Ä®¤n¤µ„„ªµ­n ¼˜¦°µ®µ¦š¸Éž¦³„°—ª¥œo Ê嘵¨¤°¨Ã˜­°¥nµŠ¤¸œ´¥­Îµ‡´ Ž¹ÉŠ‡¨oµ¥ „´„µ¦š—¨°Š °Š Ng and Saleh (2010b) š¸É«¹„¬µ„µ¦ ¥µ¥¡œ›´ »r„¨oª¥Å¤o Paphiopedilum rothschildianum ×¥­µ¤µ¦™´„œÎµÄ®oÁ„—¥°—š· ª¸‡¼–‹µ„˜µ oµŠ Ž¹ÉŠ¤¸‹Îµœªœ¥°—ÁŒ¨¸É¥ 2.9 ¥°— ®¨´Š‹µ„Á¡µ³Á¨¸Ê¥ŠÁž}œÁª¨µ 16 ­´ž—µ®r ‹µ„­¼˜¦°µ®µ¦ 1/2 MS š¸Éž¦³„°—ª¥ÁžžÃ˜œ‡ªµ¤o Á o¤ oœ 1.0 „¦´¤˜n°¨·˜¦ œÊ嘵¨Ž¼Ã‡¦­ ¨³Å¤n¤¸­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ ˜n˜nµŠ‹µ„„µ¦š—¨°Š °Š Chen et al. (2002) š¸É«¹„¬µ„µ¦Á„·—¥°—šª¸‡¼–‹µ„˜µ °Š„oµœn°—°„„¨oª¥Å¤ o Paphiopedilum philippinense 2 ­µ¥¡œ›´ »r (PH59 ¨³ PH60) Ĝ­µ¥¡œ›´ »r PH59 ¤¸„µ¦Á„·—¥°—¨³‹Îµœªœ¥°—˜n°

·ÊœÁœºÊ°Á¥ºÉ°¤µ„š¸É­»— 66.7 ¨³ 1.5 Áž°¦ rÁŽÈœ˜r ˜µ¤¨Îµ—´ ‹µ„­¼˜¦°µ®µ¦š¸Éž¦³„°—oª¥­µ¦‡ª‡»¤ „µ¦Á˜·Ã˜ 2,4-D ‡ªµ¤Á o¤ oœ 4.52 ŤÇ¦Ã¤¨µ¦r ¦nª¤„´ TDZ ‡ªµ¤Á o¤ oœ 0.45 ŤÇ¦Ã¤¨µ¦r ¨³­µ¥¡œ›´ »r PH60 ¤¸„µ¦Á„·—¥°—¤µ„š¸É­»— 80 Áž°¦rÁŽÈœ˜r ‹µ„­¼˜¦°µ®µ¦š¸Éž¦³„°—ª¥­µ¦o ‡ª‡»¤„µ¦Á˜Ã˜· 2,4-D ‡ªµ¤Á o¤ oœ 4.52 ŤÇ¦Ã¤¨µ¦r ¦nª¤„´ TDZ ‡ªµ¤Á o¤ oœ 0.45 ŤÇ¦ 䨵¦r

5. «¹„¬µ­¼˜¦°µ®µ¦š¸ÉÁ®¤µ³­¤˜n°„µ¦´„œÎµ„µ¦Á„·—‡¨¨´­ ®¦º° somatic embryos ‹µ„·ÊœÁœºÊ° Á¥ºÉ°¦³®ªnµŠ o° °Š„¨oª¥Å¤ o Paphiopedilum ‘Delrosi’ ‹µ„„µ¦œÎµœÁœ·Ê ºÊ°Á¥ºÉ°¦³®ªnµŠ o° °Š„¨oª¥Å¤o Paphiopedilum ‘Delrosi’ ŞÁ¡µ³Á¨¥Š¸Ê œ­¼˜¦°µ®µ¦ 1/2 MS š¸Éž¦³„°—ª¥œo Ê嘵¨ 3 œ·— ‡º° Ž¼Ã‡¦­ „¨¼Ã‡­ ¨³¤°¨Ã˜­ ‡ªµ¤ Á o¤ oœ 20 „¦´¤˜n°¨·˜¦ ¦nª¤„´­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ TDZ ‡ªµ¤Á o¤ oœ 0.45 ¨³ 4.54 ŤÇ¦Ã¤ ¨µ¦r Á¡¸¥Š°¥nµŠÁ—¸¥ª ®¦º°¦nª¤„´ 2,4-D ‡ªµ¤Á o¤ oœ 4.52 ¨³ 13.56 ŤÇ¦Ã¤¨µ¦r Áž}œ¦³¥³Áª¨µ 16 ­´ž—µ®r ¡ªnµ ·ÊœÁœ°Á¥ºÊ ºÉ°¦³®ªnµŠ °šo ¸ÉÁ¡µ³Á¨¸Ê¥Šœš»„­¼˜¦°µ®µ¦ oµŠ˜oœÅ¤n¤¸„µ¦Á„·—‡¨¨´­ 79

®¦º° somatic embryos Á¨¥ ÁœºÉ°Š‹µ„œÁœ·Ê ºÊ°Á¥ºÉ°¦³®ªnµŠ o°¤¸„µ¦Áž¨¸É¥œÂž¨ŠÁžœ­} ¸œÊ嘵¨Â¨³˜µ¥ ®¨´Š‹µ„Á¦·É¤Á¡µ³Á¨¸Ê¥ŠÁžœ¦³¥³Áª¨µž¦³¤µ–} 4-5 ­´ž—µ® r Ž¹ÉŠÂ˜„˜nµŠ„´„µ¦š—¨°Š °Š Lin (1986) š¸É«¹„¬µ„µ¦ ¥µ¥¡œ›´ »r„¨oª¥Å¤o Phalaenopsis ¨³ Doritaenopsis ‹µ„„µ¦Á¡µ³Á¨¸Ê¥ŠÁœºÊ°Á¥ºÉ° ¦³®ªnµŠ o° °Š„oµœn°—°„ ×¥­µ¤µ¦™´„œÎµÄ®oÁ„·— PLBs ‹µ„ÁœºÊ°Á¥ºÉ°­nªœÁ°¡·Á—°¦r¤·­ ¨³ Ÿ·ª®œoµ °Š¦°¥˜´— ¨³¡´•œµÁž}œ˜oœš¸É­¤¼¦–r®¨´Š‹µ„Á¡µ³Á¨¸Ê¥ŠÁžœÁª¨µ} 60 ª´œ œ­¼˜¦°µ®µ¦ VW š¸Éž¦³„°—oª¥­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ BA ‡ªµ¤Á ¤ o oœ 1 ¨³ 5 ¤·¨¨·„¦´¤˜n°¨·˜¦ ¨³„µ¦ š—¨°Š °Š Chen et al. (2002) š¸É«¹„¬µ„µ¦ ¥µ¥¡œ›´ »r„¨oª¥Å¤ o Epidendrum radicans ‹µ„„µ¦ Á¡µ³Á¨¸Ê¥ŠÁœºÊ°Á¥ºÉ°¦³®ªnµŠ o° °Š„oµœn°—°„ ×¥­µ¤µ¦™´„œÎµ„µ¦Á„·— PLBs œ­¼˜¦°µ®µ¦ 1/2 MS š¸Éž¦³„°—oª¥­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ TDZ ‡ªµ¤Á o¤ oœ 0.45 ŤÇ¦Ã¤¨µ¦r ĜÁª¨µ 2 Á—º°œ ¨³­µ¤µ¦™Á¡·É¤‹Îµœªœ PLBs ŗo¤µ„š¸É­»— 23.6 PLBs Ĝ­¼˜¦°µ®µ¦š¸Éž¦³„°—ª¥œo Ê嘵¨Ž¼Ã‡¦­

‡ªµ¤Á o¤ oœ 40 „¦´¤˜n°¨·˜¦ NH4NO3 ‡ªµ¤Á o¤ oœ 825 ¤·¨¨·„¦´¤˜n°¨·˜¦ ¨³ KNO3 ‡ªµ¤Á o¤ oœ 950 ¤·¨¨·„¦´¤˜n°¨·˜¦ ¨³ PLBs ­µ¤µ¦™¡´•œµÁž}œ˜oœš¸É­¤¼¦–rÁ¤ºÉ°Á¡µ³Á¨¸Ê¥Šœ­¼˜¦°µ®µ¦š¸É

ž¦³„°—ª¥­µ¦‡ª‡o »¤„µ¦Á˜·Ã˜ TDZ š¸É‡ªµ¤Á o¤ oœÁ—·¤ ĜÁª¨µ 1 Á—º°œ

6. «¹„¬µ­¼˜¦°µ®µ¦š¸ÉÁ®¤µ³­¤˜n°„µ¦´„œÎµ„µ¦Á„·—‡¨¨´­ ®¦º° somatic embryos ‹µ„·Êœ­nªœÄ °Š„¨oª¥Å¤ o Paphiopedilum ‘Delrosi’

‹µ„„µ¦œÎµÄ¦·Áª–¥°—‹µ„˜oœ„¨ª¥Å¤o o Paphiopedilum ‘Delrosi’ ‡¦¹ÉŠ¨nµŠ œµ— 0.3 x 0.4 ˜µ¦µŠÁŽœ˜·Á¤˜¦ ŞÁ¡µ³Á¨¸Ê¥Šœ­ ¼˜¦°µ®µ¦ 1/2 MS š¸Éž¦³„°—ª¥œo Ê嘵¨ 3 œ·— ‡º° Ž¼Ã‡¦­ „¨¼Ã‡­ ¨³¤°¨Ã˜­ ‡ªµ¤Á o¤ oœ 20 „¦´¤˜°¨n ·˜¦ ¦nª¤„´­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ TDZ ‡ªµ¤Á o¤ oœ 0.45 ¨³ 4.54 ŤÇ¦Ã¤¨µ¦r Á¡¸¥Š°¥nµŠÁ—¸¥ª ®¦º°¦nª¤„´ 2,4-D ‡ªµ¤Á o¤ oœ 4.52 ¨³ 13.56 ŤÇ¦Ã¤¨µ¦r Áž}œ¦³¥³Áª¨µ 16 ­´ž—µ®r ¡ªnµ ·Êœ­nªœÄš¸ÉÁ¡µ³Á¨¸Ê¥Šœš»„­¼˜¦°µ®µ¦ oµŠ˜oœÅ¤n¤¸„µ¦Á„·—‡¨¨´­ ®¦º° somatic embryos Á¨¥ ÁœºÉ°Š‹µ„·Êœ­ªœÄ¤n „µ¦Áž¨¸ ¸É¥œÂž¨ŠÁž}œ­¸ œÊ嘵¨Â¨³˜µ¥®¨´Š‹µ„Á¦·É¤Á¡µ³Á¨¸Ê¥ŠÁžœ¦³¥³Áª¨µž¦³¤µ–} 1-2 ­´ž—µ®r Ž¹ÉŠÂ˜„˜nµŠ„´„µ¦š—¨°Š °Š Chen et al. (2004) š¸É«¹„¬µ„µ¦ ¥µ¥¡´œ›»r„¨oª¥Å¤o Paphiopedilum hybrid 2 ­µ¥¡´œ›»r (PH59 ¨³ PH60) ‹µ„„µ¦´„œÎµÄ®oÁ„·—¥°—‹µ„¦·Áª–µ—Ÿ¨ °Š·Êœ­nªœÄ š¸É­µ¤µ¦™Á¡·É¤‹Îµœªœ¥°— ÁŒ¨¸É¥˜n°·Êœ­nªœÄš¸Éčoš—¨°ŠÅ—o—¸š¸É­»— œ­¼˜¦°µ®µ¦š¸Éž¦³„°—ª¥­µ¦‡ª‡o »¤„µ¦Á˜·Ã˜ TDZ ‡ªµ¤Á o¤ oœ 4.54 ŤÇ¦Ã¤¨µ¦r Ĝ­µ¥¡´œ›»r PH59 ¨³­µ¤µ¦™´„œÎµÄ®oÁ„·—¥°—‹µ„·Êœ­nªœÄš¸É čoš—¨°ŠÅ—o—¸š¸É­»— œ­¼˜¦°µ®µ¦š¸Éž¦³„°—oª¥­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ 2,4-D ‡ªµ¤Á o¤ oœ 4.52 ŤÇ¦Ã¤¨µ¦r ¦nª¤„´ TDZ 0.45 ŤÇ¦Ã¤¨µ¦r Ĝ­µ¥¡´œ›»r PH60 ¨³„µ¦š—¨°Š °Š Chen and Chang (2006) š¸É«¹„¬µ„µ¦ ¥µ¥¡œ›´ »r„¨oª¥Å¤o Phalaenopsis amabilis ‹µ„·Êœ­nªœÄ Ž¹ÉŠ­µ¤µ¦™´„ 80

œÎµÄ®oÁ„·— somatic embryos ¤µ„š¸É­»—ÁŒ¨¸É¥ 19.4 ‹µ„ epidermal cell ×¥˜¦Š œ­¼˜¦­¼˜¦°µ®µ¦ 1/2 MS š¸Éž¦³„°—oª¥­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ TDZ ‡ªµ¤Á o¤ oœ 3.0 ŤÇ¦Ã¤¨µ¦r ¨³„µ¦š—¨°Š °Š Park et al. (2002b) š¸É«¹„¬µ„µ¦ ¥µ¥¡´œ›»r„¨oª¥Å¤o Doritaenopsis hybrid ‹µ„·Êœ­nªœÄ—ª¥o ª·›¸ thin-section culture Á¡ºÉ°´„œÎµÄ®oÁ„·— PLBs ×¥˜¦Š Ž¹ÉŠ­µ¤µ¦™´„œÎµÄ®oÁ„·— PLBs ŗo 72.3 Áž°¦rÁŽÈœ˜r ¨³¤¸‹Îµœªœ PLBs ÁŒ¨¸É¥ 18 PLBs œ­¼˜¦°µ®µ¦ 1/2 MS š¸Éž¦³„°—ª¥­µ¦‡ª‡o »¤ „µ¦Á˜·Ã˜ TDZ ‡ªµ¤Á o¤ oœ 9.0 ŤÇ¦Ã¤¨µ¦r

šš¸É 6

­¦»žŸ¨„µ¦š—¨°Š

1. «¹„¬µ­¼˜¦°µ®µ¦š¸ÉÁ®¤µ³­¤˜n°„µ¦´„œÎµ„µ¦Á„·—¥°—šª¸‡¼–‹µ„­ªœ¥°— °Š„¨n oª¥Å¤ o Paphiopedilum ‘Delrosi’ 1.1 «¹„¬µŸ¨ °Š­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ TDZ, BA, Kn, 2,4-D ¨³ Ads ˜n°„µ¦´„œÎµ„µ¦Á„·—¥°— šª¸‡¼–‹µ„­nªœ¥°— °Š„¨oª¥Å¤ o Paphiopedilum ‘Delrosi’ „µ¦´„œÎµ„µ¦Á„·—¥°—šª¸‡¼–‹µ„­nªœ¥°— °Š„¨oª¥Å¤o Paphiopedilum ‘Delrosi’ ×¥

Á¡µ³Á¨¸Ê¥Šœ­¼˜¦°µ®µ¦—´—ž¨Š Hyponex [ž»l¥ Hyponex (6.5N-4.5P-19K) ‡ªµ¤Á o¤ oœ 3 „¦´¤ ˜n°¨·˜¦ + ÁžžÃ˜œ (peptone) ‡ªµ¤Á o¤ oœ 2 „¦´¤˜n°¨·˜¦ + œÊε˜o¤¤´œ ¦´ÉŠ 100 „¦´¤˜°¨n ·˜¦] ¦nª¤„´

­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ TDZ, BA, Kn, 2,4-D ¨³ Ads ‡ªµ¤Á o¤ oœ˜nµŠÇ Áž}œ¦³¥³Áª¨µ 16 ­´ž—µ® r ¡ªnµ ­¼˜¦°µ®µ¦—´—ž¨Š Hyponex š¸Éž¦³„°—ª¥­µ¦‡ª‡o »¤„µ¦Á˜·Ã˜ Kn ‡ªµ¤Á o¤ oœ 4.65

ŤÇ¦Ã¤¨µ¦r ¦nª¤„´ 2,4-D ‡ªµ¤Á o¤ oœ 4.52 ŤÇ¦Ã¤¨µ¦r Ä®oÁž°¦rÁŽÈœ˜r„µ¦Á„—¥°—ÁŒ¨· ¸É¥¤µ„ š¸É­»— 100 Áž°¦rÁŽÈœ˜r ¨³Ä®o‹Îµœªœ¥°—Ä®¤nÁŒ¨¸É¥¤µ„¨³­¤ÉεÁ­¤°š¸É­»— 1.4 ¥°—

1.2 «¹„¬µŸ¨ °Š­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ TDZ BA Kn ¨³ 2,4-D ˜n°„µ¦´„œÎµ„µ¦Á„·—¥°—šª¸‡¼– ‹µ„˜oœ„¨oª¥Å¤o Paphiopedilum ‘Delrosi’ „µ¦´„œÎµ„µ¦Á„·—¥°—šª¸‡¼–‹µ„­nªœ¥°— °Š„¨oª¥Å¤o Paphiopedilum ‘Delrosi’ ×¥ Á¡µ³Á¨¸Ê¥Šœ­¼˜¦°µ®µ¦—´—ž¨Š Hyponex [ž»l¥ Hyponex (6.5N-4.5P-19K) ‡ªµ¤Á o¤ oœ 3 „¦´¤ ˜n°¨·˜¦ + ÁžžÃ˜œ (peptone) ‡ªµ¤Á o¤ oœ 2 „¦´¤˜n°¨·˜¦ + œÊε˜o¤¤´œ ¦´ÉŠ 100 „¦´¤˜°¨n ·˜¦] ¦nª¤„´ ­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ TDZ, BA, Kn ¨³ 2,4-D ‡ªµ¤Á o¤ oœ˜nµŠÇ Áž}œ¦³¥³Áª¨µ 16 ­´ž—µ® r ¡ªnµ ­¼˜¦°µ®µ¦—´—ž¨Š Hyponex š¸Éž¦³„°—ª¥­µ¦‡ª‡o »¤„µ¦Á˜·Ã˜ TDZ ‡ªµ¤Á o¤ oœ 0.45 ŤÇ¦Ã¤¨µ¦r ¦nª¤„´ 2,4-D ‡ªµ¤Á o¤ oœ 4.52 ŤÇ¦Ã¤¨µ¦r Ä®oÁž°¦rÁŽÈœ˜r„µ¦Á„—¥°—Ä®¤· nÁŒ¨¸É¥ ¤µ„š¸É­»— 90 Áž°¦rÁŽÈœ˜r ¨³Ä®o‹Îµœªœ¥°—Ä®¤nÁŒ¨¥¤µ„¨³­¤¸É ÉεÁ­¤°š¸É­»— 1.3 ¥°—

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2. «¹„¬µœ·— °ŠœÊ嘵¨š¸ÉÁ®¤µ³­¤˜n°„µ¦´„œÎµ„µ¦Á„·—¥°—šª¸‡¼–‹µ„­nªœ¥°— °Š„¨oª¥Å¤o Paphiopedilum ‘Delrosi’ „µ¦´„œÎµ„µ¦Á„·—¥°—šª¸‡¼–‹µ„­nªœ¥°— °Š„¨oª¥Å¤o Paphiopedilum ‘Delrosi’ ×¥ Á¡µ³Á¨¸Ê¥Šœ­¼˜¦°µ®µ¦—´—ž¨Š Hyponex [ž»l¥ Hyponex (6.5N-4.5P-19K) ‡ªµ¤Á ¤ o oœ 3 „¦´¤˜n° ¨·˜¦ + ÁžžÃ˜œ (peptone) ‡ªµ¤Á o¤ oœ 2 „¦´¤˜n°¨·˜¦ + œÊε˜o¤¤´œ ¦´ÉŠ 100 „¦´¤˜n°¨·˜¦] š¸É ž¦³„°—ª¥œo Ê嘵¨ 3 œ·— ‡º° Ž¼Ã‡¦­ „¨¼Ã‡­ ¨³¤°¨Ã˜­ ‡ªµ¤Á o¤ oœ 20 „¦´¤˜n°¨·˜¦ ¦nª¤„´ ­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ TDZ ‡ªµ¤Á o¤ oœ 0.45 ŤÇ¦Ã¤¨µ¦r ¦nª¤„´ 2,4-D ‡ªµ¤Á o¤ oœ 4.52 Ť Ǧ䨵¦r ®¦º° Kn ‡ªµ¤Á o¤ oœ 4.65 ŤÇ¦Ã¤¨µ¦r ¦nª¤„´ 2,4-D ‡ªµ¤Á o¤ oœ 4.52 ŤÇ¦Ã¤ ¨µ¦r Áž}œ¦³¥³Áª¨µ 16 ­´ž—µ®r ¡ªnµ ­¼˜¦°µ®µ¦—´—ž¨Š Hyponex š¸Éž¦³„°—ª¥œo Ê嘵¨„¨¼Ã‡­ ‡ªµ¤Á o¤ oœ 20 „¦´¤˜n°¨·˜¦ ¨³­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ TDZ ‡ªµ¤Á o¤ oœ 0.45 ŤÇ¦Ã¤¨µ¦r ¦nª¤„´ 2,4-D ‡ªµ¤Á o¤ oœ 4.52 ŤÇ¦Ã¤¨µ¦r Ä®oÁž°¦rÁŽÈœ˜r„µ¦Á„—¥°—Ä®¤· nÁŒ¨¥¤µ„š¸É ¸É­»— 80 Áž°¦rÁŽÈœ˜r ¨³Ä®o‹Îµœªœ¥°—Ä®¤nÁŒ¨¸É¥ 1.8 ¥°—

3. «¹„¬µ­¼˜¦°µ®µ¦š¸ÉÁ®¤µ³­¤˜n°„µ¦´„œÎµ„µ¦Á„·—¥°—šª¸‡¼–‹µ„˜œ„¨o oª¥Å¤o Paphiopedilum ‘Delrosi’ š¸É˜´—­nªœ¥°—°°„ „µ¦´„œÎµ„µ¦Á„·—¥°—šª¸‡¼–‹µ„˜oœ„¨oª¥Å¤o Paphiopedilum ‘Delrosi’ š¸É˜´—¥°—°°„

×¥Á¡µ³Á¨¸Ê¥Šœ­¼˜¦°µ®µ¦—´—ž¨Š Hyponex [ž»l¥ Hyponex (6.5N-4.5P-19K) ‡ªµ¤Á o¤ oœ 3

„¦´¤˜n°¨·˜¦ + ÁžžÃ˜œ (Peptone) ‡ªµ¤Á o¤ oœ 2 „¦´¤˜n°¨·˜¦ + œÊ嘤¤o ´œ ¦´ÉŠ 100 „¦´¤˜n°¨·˜¦] š¸É ž¦³„°—ª¥œo Ê嘵¨ 3 œ·— ‡º° Ž¼Ã‡¦­ „¨¼Ã‡­ ¨³¤°¨Ã˜­ ‡ªµ¤Á o¤ oœ 20 „¦´¤˜n°¨·˜¦ ¦nª¤„´ ­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ TDZ ‡ªµ¤Á o¤ oœ 0.45 ŤÇ¦Ã¤¨µ¦r ®¦º° TDZ ‡ªµ¤Á o¤ oœ 0.45 ŤÇ¦ 䨵¦r ¦nª¤„´ 2,4-D ‡ªµ¤Á o¤ oœ 4.52 ŤÇ¦Ã¤¨µ¦r ®¦º° Kn ‡ªµ¤Á o¤ oœ 4.65 ŤÇ¦Ã¤¨µ¦r ¦nª¤„´ 2,4-D ‡ªµ¤Á o¤ oœ 4.52 ŤÇ¦Ã¤¨µ¦r Áž}œ¦³¥³Áª¨µ 16 ­´ž—µ®r ¡ªnµ ­¼˜¦°µ®µ¦ —´—ž¨Š Hyponex š¸Éž¦³„°—oª¥œÊ嘵¨Ž¼Ã‡¦­ ‡ªµ¤Á o¤ oœ 20 „¦´¤˜n°¨·˜¦ ¨³­µ¦‡ª‡»¤„µ¦ Á˜·Ã˜ Kn ‡ªµ¤Á o¤ oœ 4.65 ŤÇ¦Ã¤¨µ¦r ¦nª¤„´ 2,4-D ‡ªµ¤Á o¤ oœ 4.52 ŤÇ¦Ã¤¨µ¦r Ä®o Áž°¦rÁŽÈœ˜r„µ¦Á„·—¥°—Ä®¤nÁŒ¨¸É¥—¸š¸É­»— 90 Áž°¦rÁŽÈœ˜r ¨³Ä®o‹Îµœªœ¥°—Ä®¤nÁŒ¨¸É¥ 1.6 ¥°—

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4. «¹„¬µ­¼˜¦°µ®µ¦š¸ÉÁ®¤µ³­¤˜n°„µ¦´„œÎµ„µ¦Á„·—¥°—‹µ„­nªœ o° °Š˜œ„¨o oª¥Å¤o Paphiopedilum ‘Delrosi’ „µ¦´„œÎµ„µ¦Á„·—¥°—‹µ„­ªœ n o° °Š„¨oª¥Å¤o Paphiopedilum ‘Delrosi’ ×¥ Á¡µ³Á¨¸Ê¥Šœ­¼˜¦°µ®µ¦—´—ž¨Š Hyponex [ž»l¥ Hyponex (6.5N-4.5P-19K) ‡ªµ¤Á o¤ oœ 3 „¦´¤ ˜n°¨·˜¦ + ÁžžÃ˜œ (Peptone) ‡ªµ¤Á o¤ oœ 2 „¦´¤˜n°¨·˜¦ + œÊε˜o¤¤´œ ¦´ÉŠ 100 „¦´¤˜n°¨·˜¦] š¸É ž¦³„°—ª¥œo Ê嘵¨ 3 œ·— ‡º° Ž¼Ã‡¦­ „¨¼Ã‡­ ¨³¤°¨Ã˜­ ‡ªµ¤Á o¤ oœ 20 „¦´¤˜n°¨·˜¦ ¦nª¤„´ ­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ TDZ ‡ªµ¤Á o¤ oœ 0.45 ŤÇ¦Ã¤¨µ¦r ®¦º° TDZ ‡ªµ¤Á o¤ oœ 0.45 ŤÇ¦ 䨵¦r ¦nª¤„´ 2,4-D ‡ªµ¤Á o¤ oœ 4.52 ŤÇ¦Ã¤¨µ¦r ®¦º° Kn ‡ªµ¤Á o¤ oœ 4.65 ŤÇ¦Ã¤¨µ¦r ¦nª¤„´ 2,4-D ‡ªµ¤Á o¤ oœ 4.52 ŤÇ¦Ã¤¨µ¦r Áž}œ¦³¥³Áª¨µ 16 ­´ž—µ®r ¡ªnµ ­¼˜¦°µ®µ¦ —´—ž¨Š Hyponex š¸Éž¦³„°—oª¥œÊ嘵¨Ž¼Ã‡¦­ ‡ªµ¤Á o¤ oœ 20 „¦´¤˜n°¨·˜¦ š¸Éž¦µ«‹µ„­µ¦ ‡ª‡»¤„µ¦Á˜Ã˜· Ä®oÁž°¦rÁŽÈœ˜r„µ¦Á„·—¥°—ÁŒ¨¸É¥ 100 Áž°¦rÁŽÈœ˜r ¨³Ä®o‹Îµœªœ¥°—Ä®¤nÁŒ¨¸É¥ 1.0 ¥°—

5. «¹„¬µ­¼˜¦°µ®µ¦š¸ÉÁ®¤µ³­¤˜n°„µ¦´„œÎµ„µ¦Á„·—‡¨¨´­ ®¦º° somatic embryos ‹µ„·ÊœÁœºÊ° Á¥ºÉ°¦³®ªnµŠ o° °Š„¨oª¥Å¤ o Paphiopedilum ‘Delrosi’ „µ¦´„œÎµ„µ¦Á„·—‡¨¨´­ ®¦º° somatic embryos ‹µ„·ÊœÁœºÊ°Á¥ºÉ°¦³®ªnµŠ o° °Š

„¨oª¥Å¤ o Paphiopedilum ‘Delrosi’ ×¥Á¡µ³Á¨¸Ê¥Šœ­¼˜¦°µ®µ¦ 1/2 MS (1/2 strength macro-and full-strength micro elements) š¸Éž¦³„°— oª¥œÊ嘵¨ 3 œ·— ‡º° Ž¼Ã‡¦­ „¨¼Ã‡­ ¨³¤°¨Ã˜­ ‡ªµ¤Á o¤ oœ 20 „¦´¤˜n°¨·˜¦ ¦nª¤„´­µ¦‡ª‡»¤„µ¦Á˜Ã˜· TDZ ‡ªµ¤Á o¤ oœ 0.45 ¨³ 4.54 Ť Ǧ䨵¦r Á¡¸¥Š°¥nµŠÁ—¸¥ª ®¦º°¦nª¤„´ 2,4-D ‡ªµ¤Á ¤ o oœ 4.52 ¨³ 13.56 ŤÇ¦Ã¤¨µ¦r Áž}œ ¦³¥³Áª¨µ 16 ­´ž—µ®r ¡ªnµ ·ÊœÁœºÊ°Á¥ºÉ°¦³®ªnµŠ o°š¸ÉÁ¡µ³Á¨¸Ê¥Šœš»„­¼˜¦°µ®µ¦ oµŠ˜oœÅ¤n¤¸„µ¦Á„·— ‡¨¨´­ ®¦º° somatic embryos Á¨¥ ÁœºÉ°Š‹µ„·ÊœÁœºÊ°Á¥ºÉ°¦³®ªnµŠ o°¤„µ¦Áž¨¸ ¸É¥œÂž¨ŠÁž}œ­¸œÊ嘵¨ ¨³˜µ¥®¨´Š‹µ„Á¦·É¤Á¡µ³Á¨¸Ê¥ŠÁž}œ¦³¥³Áª¨µž¦³¤µ– 4-5 ­´ž—µ® r

6. «¹„¬µ­¼˜¦°µ®µ¦š¸ÉÁ®¤µ³­¤˜n°„µ¦´„œÎµ„µ¦Á„·—‡¨¨´­ ®¦º° somatic embryos ‹µ„·Êœ­nªœÄ °Š„¨oª¥Å¤ o Paphiopedilum ‘Delrosi’ „µ¦´„œÎµ„µ¦Á„·—‡¨¨´­ ®¦º° somatic embryos ‹µ„·Êœ­ªœÄ °Š„¨n oª¥Å¤o Paphiopedilum ‘Delrosi’ ×¥Á¡µ³Á¨¸Ê¥Šœ­¼˜¦°µ®µ¦ 1/2 MS (1/2 strength macro-and full- strength micro elements) š¸Éž¦³„°—ª¥œo Ê嘵¨ 3 œ·— ‡º° Ž¼Ã‡¦­ „¨¼Ã‡­ ¨³¤°¨Ã˜­ ‡ªµ¤ Á o¤ oœ 20 „¦´¤˜n°¨·˜¦ ¦nª¤„´­µ¦‡ª‡»¤„µ¦Á˜·Ã˜ TDZ ‡ªµ¤Á o¤ oœ 0.45 ¨³ 4.54 ŤÇ¦Ã¤ 84

¨µ¦r Á¡¸¥Š°¥nµŠÁ—¸¥ª ®¦º°¦nª¤„´ 2,4-D ‡ªµ¤Á o¤ oœ 4.52 ¨³ 13.56 ŤÇ¦Ã¤¨µ¦r Áž}œ¦³¥³Áª¨µ 16 ­´ž—µ®r ¡ªnµ ·Êœ­nªœÄš¸ÉÁ¡µ³Á¨¸Ê¥Šœš»„­¼˜¦°µ®µ¦ oµŠ˜oœÅ¤n¤¸„µ¦Á„·—‡¨¨´­ ®¦º° somatic embryos Á¨¥ ÁœºÉ°Š‹µ„œ­·Ê nªœÄ¤„µ¦Áž¨¸ ¸É¥œÂž¨ŠÁž}œ­¸œÊ嘵¨Â¨³˜µ¥®¨´Š‹µ„Á¦·É¤Á¡µ³Á¨¸Ê¥ŠÁž}œ ¦³¥³Áª¨µž¦³¤µ– 1-2 ­´ž—µ®r

85

ขอเสนอแนะ 1. การนําตนกลวยไมไวในท ี่มืดเพื่อใหเนอเยื้ ื่อระหวางขอ และใบยืดยาวพอที่จะใชทํา การทดลองควรใชสารควบคุมการเติบโตในกลุมที่ชวยใหเซลลยืดตัวไดดีรวมดวยเพอลดระยะเวลาื่ ของขั้นตอนการเตรียมพืช 2. การชักนําการเกิดยอดทวีคูณจากกลวยไม Paphiopedilum ‘Delrosi’ สวนตางๆ ควร นํายอดใหมไปปลูกและหาอัตราการรอดชีวิตจากยอดที่ไดจากสวนต างๆ 3. การชักนําการเกิดยอดทวีคูณจากกลวยไม Paphiopedilum ‘Delrosi’ สวนตางๆ ควร ศึกษาตอไปว ายอดใหมที่ไดจากส วนตางๆ มีลักษณะเหมอนตื นเดิมหรอแตกตื างกนอยั างไร 4. การเพาเลี้ยงสวนเนื้อเยื่อระหวางขอ และชิ้นสวนใบ ควรใชภาชนะเพาะเลี้ยงใหมี ขนาดเหมาะสมกับเนื้อเยื่อ เพื่อลดการสูญเสียน้ําของชิ้นเนื้อเยื่อ

86

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£µ¬µÅš¥ „¦³š¦ªŠÁ„¬˜¦Â¨³­®„¦–r. nµªž¦³µ­´¤¡´œ›.r [Online] Accessed 07 April 2007. Available from http://www.moac.go.th/builder/moac02/information/view_index.php?id=2489. ______. nµªž¦³µ­´¤¡œ›´ .r [Online] Accessed 01 February 2010. Available from http://webnew.moac.go.th/ewt_news.php?nid=991&filename=index. ______. nµªž¦³µ­´¤¡œ›´ .r [Online] Accessed 16 December 2010. Available from http://webnew.moac.go.th/ewt_news.php?nid=4683&filename=index. ‡Îµœ¼œ „µ‹œ£¼¤·. „µ¦Á¡µ³Á¨¸Ê¥ŠÁœºÊ°Á¥ºÉ°¡º. ¡·¤¡r‡¦´ÊŠš¸É 2. „¦»ŠÁš¡² : ­Îµœ´„¡·¤¡r®nŠ‹»¯µ¨Š„¦–r ¤®µª·š¥µ¨´¥, 2544. ›µ¦š·¡¥r Á¡¦³¼¦–·œ. Ÿ¨·Ä. [Online] Accessed 27 January 2010. Available from

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›»¦„·‹Ã¦Š¡·¤¡r ¦·¬´š°¤¦·œš¦r¡¦·Êœ˜·ÊŠÂ°œ—r¡´¨··ÉŠ ‹Îµ„´— (¤®µœ), 2549.

¦³¡¸ ­µ‡¦·„. „¨oª¥Å¤o¦°ŠÁšoµœµ¦¸ ª·›¸ž¨ „Á¨¼ ¸Ê¥ŠÂ¨³ž{®µ°œ»¦´„¬r›¦¦¤µ˜.· „¦»ŠÁš¡² : ­Îµœ´„¡·¤¡r ðÁ—¸¥œ­Ã˜¦r, 2535. ______. „µ¦Á¡µ³ž¨¼„„¨oª¥Å¤Äœ­£µ¡Âª—¨o o°¤ °Šž¦³Áš«Åš¥. „¦»ŠÁš¡² : ªœ¡·¤¡r, 2516 ¦´Š­§¬—·Í „µª¸˜p³. „µ¦Á¡µ³Á¨¸Ê¥ŠÁœºÊ°Á¥ºÉ°¡º : ®¨´„„µ¦Â¨³Áš‡œ·‡. ¡·¤¡r‡¦´ÊŠš¸É 2 „¦»ŠÁš¡² : ­Îµœ´„¡·¤¡r¤®µª·š¥µ¨´¥Á„¬˜¦«µ­˜¦r, 2541. °»Å¦ ‹·¦¤Š‡¨„µ¦. „¨oª¥Å¤¦°ŠÁšo oµœµ¦¸. ¡·¤¡r‡¦´ÊŠš¸É 4. „¦»ŠÁš¡² : ­Îµœ´„¡·¤¡roµœÂ¨³­ªœ ­µ¥ ›»¦„·‹Ã¦Š¡·¤¡r ¦·¬´š°¤¦·œš¦r¡¦·Êœ˜·ÊŠÂ°œ—r¡´¨··ÉŠ ‹Îµ„´— (¤®µœ), 2549.

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92

คําอธิบายคายํ อ คํายอ คําเต็ม μM = Micromolar 2,4-D = 2,4-dichlorophenoxyacetic acid Ads = Adenine sulfate BA = 6-benzyladenine g/l = Gram/litre Kn = Kinetin mg/l = Milligram/litre MS = Murashige and Skoog (1962) TDZ = Thidiazuron v/v = Volume per volume

93

ตารางภาคผนวก องคประกอบของอาหารสูตร Murashige and Skoog (1962)

องคประกอบ มิลลิกรัมตอลิตร ธาตุอาหารหลัก (macronutrients)

MgSO4.7H2O 370

CaCl2.2H2O 440

KNO3 1,900

NH4NO3 1,650

KHPO4 170 ธาตุอาหารรอง (micronutrients)

FeSO4.7H2O 27.8

MnSO4.4H2O 22.3

KI 0.83

CoCl2.6H2O 0.025

ZnSO4.5H2O 8.6

CuSO4.5H2O 0.025

H3BO3 6.2

Na2MoO4.2H2O 0.25 สารอินทรีย (organic constituents) Glycine 2 Myo-inositol 100 Thiamine HCl (Vitamin B1) 0.1 Pyridoxine HCl (Vitamin B2) 0.5 Nicotinic acid 0.5 EDTA (disodium salt) 37.3 Sucrose 30,000 (30 g/l) Agar 6,200 (6.2 g/l)

94

ประวัติผูวิจัย

ชื่อ-สกุล นายเอกสิทธิ์ นิสยันต  ที่อยู 61/2 หมู 1 ตําบล เขาพระ อําเภอ เดิมบางนางบวช จังหวัด สุพรรณบุรี 72120 ประวัติการศึกษา พ.ศ. 2545 สําเร็จการศึกษาชั้นมัธยมศกษาปึ ที่ 6 จากโรงเรียนกาญจนาภิเษกวทยาลิ ัย สุพรรณบุรี พ.ศ. 2549 สําเร็จการศึกษาปริญญาวิทยาศาสตรบัณฑิต คณะวิทยาศาสตร สาขา ชีววิทยา จากมหาวิทยาลัยศิลปากร การศึกษารายบุคคลเรื่อง “การเพาะเลี้ยงเนอเยื้ ื่อพืชกาบหอยแครง (Dionaea muscipula)”

พ.ศ. 2550 ศึกษาตอระดบปรั ิญญามหาบัณฑิต สาขาชีววิทยา บัณฑิตวิทยาลัย มหาวิทยาลัยศลปากริ วิทยานิพนธเรองื่ “การเพาะเลี้ยงเนื้อเยื่อกลวยไมรองเทานารี (Paphiopedilum ‘Delrosi’)”

ประวัติการเขารวมอบรม

พ.ศ. 2551 เขารวมสัมมนาประจ ําปของคลัสเตอรกลวยไม ครั้งท ี่ 4 ในหวขั อ "ความกาวหนาของคล ัสเตอรกลวยไม" ณ ศูนยสงเสริมและฝกอบรม การเกษตรแหงชาต ิ มหาวิทยาลัยเกษตรศาสตร วิทยาเขตกําแพงแสน จังหวดนครปฐมั พ.ศ. 2552 เขารวมการประชุมวิชาการจลทรรศนุ แหงประเทศไทย ครั้งที่ 26 โดย ความรวมมือระหวาง มหาวิทยาลัยเชียงใหม มหาวิทยาลัยแมโจ  และ สมาคมจุลทรรศนแหงประเทศไทย พ.ศ. 2553 เสนอผลงานการวิจัยเรื่อง “Multiple shoot induction from seed derived shoots of Paphiopedilum.” The First International Orchid Symposium. National Museum of Natural Science Taichung, Taiwan. ผลงานที่ตีพิมพเผยแพร  Nisayan, E., Thepsithar, C. and Thongpukdee A. “Multiple shoot induction from seed derived shoots of Paphiopedilum.” Acta Horticulturae, 878 (2010): 199-204.