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Soil Bacteria Capable of Destroying Structure Of SOIL BACTERIA CAPABLE OF DESTROYING STRUCTURE OF CASSAVA MEALY BUGS Patcharida Khumpumuang A Thesis Submitted in Partial Fulfillment of the Requirements for the Degree of Doctor of Philosophy in Environmental Biology Suranaree University of Technology Academic Year 2015 แบคทีเรียในดินที่สามารถท าลายโครงสร้างของเพลี้ยแป้งมันส าปะหลัง นางสาวพชั ริดา คา ภูเมือง วิทยานิพนธ์นี้เป็นส่วนหนึ่งของการศึกษาตามหลักสูตรปริญญาวทิ ยาศาสตรดุษฎบี ัณฑิต สาขาวิชาชีววิทยาสิ่งแวดล้อม มหาวิทยาลัยเทคโนโลยีสุรนารี ปี การศึกษา 2558 SOIL BACTERIA CAPABLE OF DESTROYING STRUCTURE OF CASSAVA MEALY BUGS Suranaree University of Technology has approved this thesis submitted III partial fulfillment of the requirements for the Degree of Doctor of Philosophy. Chairperson ~="L..~ (Asst. Prof. Dr. Sureelak Ro tong) Member ~(Thesis A6t~dvisor) (Assoc. Prof. Dr. Jirawat Yongsawatdigul) Member 11-. tbmt' (Dr. Hathairat urai:::F!7 Member _~ca.t ~, (Assoc.~r. Kwanjai Kanokmedhakul) Member (Prof. Dr. Sukit Limpiju&e . (Prof. Dr. Santi Maensiri) Vice Rector for Academic Affairs Dean of Institute of Science and Innovation . ~ l4''If'1~1 f11mijtl~: t!ufll1~tI'hJ~'U-¥hnlJTHl'vhmtl IflHLl~l~'Utl~!l"l~tJ!!iI~lTmhtl~11~~ '" (BACTERIA IN SOIL CAPABLE OF DESTROYING STRUCTURE OF CASSAVA d'.d_~ 911 d' o:::1Q1 d' 9J MEALY BUGS) m'\lTW'VI1J'Hl'l:J1: ~'lfltlfl'lLlI'l'jl'\lTW ~'j .LlHln'I:JW 'jtl~'VItl~, 155 11'Ul. '" . 'iI )I v , !l"l~ tit! iI ~'1'111:11tlVI 'IfI ~ tI~ ~ n'U111!~ tI~'\l1n'VImll'U 'Utl ~VI'If'l'111,rVI'If!-H tlll'll tI !1J'Uuu 1:1~ . '" ~ . rYl',l"'jV" 'llfl1ftll'l tu 'Utl~lT'UftlU~ 11~~ f11'jmu f.llJ!l"l~ tit! iI ~lT'UftlU~ 11~~ I ~ tltl1rYtJ!!Ufll1 ~ tI1'U~'U JJ !J.I ?I ==< cl. I '" .:, "" & I !J.I '. I ",,1 u 'U1 I'UlJ!1J'Ul1i'VI1J'j~<tW~t!1:I~tI~tI'U mn ms fl'n'I:J1 fl H Ll'jl~!W~Lll'U 1J'j~ntlU'VI1~!fllJ'U tl~!l"l1:ltI u iI ~~!1J'Ums m11l'j 'Unau Ufll1!1 tI u 1:I~'VI~ nnu fllllJ!1J'U I U I ~'Utl~ u U fll1~ tlLllt1l4''U1.1~I'l~!ti tl n 31 I 'jJ I I '1'Unn'l'11mtl Ifl 'j ~Ll~H'U tl~!l"l~ tit! iI ~lT'UftlU ~11~~l1l"lU 1'UVI'U l1l1JJ nn 'I'111:11t1V1'1f'U'lfIlfl v I'l~ l''Utl tl ru Q tI~m .w tl 'Utl ~U 'j ~ !'VI fl'1 'VItI 'IfiJ ~ !l"l~ tIt! iI ~ff 'Ifu l"l (Phenacoccus manihoti Matile- . ~ '" Ferrero) ~~l"lU:JJ1nnl1~n 3 'lfiJ~ l1l"lufitl !l"l~tlt!iI~ff!:Utll (Phenacoccus maderiensis Green) ~ _3f r:') d d' ;f _9/ !l"l1:ltIu 1J~u '\lfl !U tI~ !1:1tI (Pseudococcus jackbeardsleyi Gimpel & Miller) u 1:1~ !l"l1:ltIu 1J~1:11tI v • ~ tFerrisia virgata Cockerel) ri'wl l'U tl~!l"l ~ tit! iI ~ff'lflJ ~ l1ffn'I:J1 JJ'If'U~h~!~ 1:1'1'U1 nl 'j ~:JJ1W 3-4 IlJ I m!lJ I'l'j JJ'lftl~!U ~ ~ u U fll1!1 tlLlllJ n flU n 'j n I~Ufi 'lftl ~U'1!dW ~ l'U U 'U'Utl~-ffl'U,r It!1:I ~ • • ril'U'ti'lt1ri'Wll (Ostioles) 'lftl~'Utl~~tllJLl~l~ 1'U~l"lUm~'\llt1mJil IUl'lllJ~U~ri'Wll 'lftl~!iJ~u'1!dW '" • !J.I 'l.c! 9J' d' Lll'U'VItl~ (Circulus) t!1:I~J11lt1 '\l (Spiracles) lJ!Ll'U~11!l''Utln1:lH 2-592-7 17-20 t!1:I~19-30 'J/ jI v iI IlJ Im!lJl'l'j mlJri'liu ~l!l"l~tlt!iI~JJm:JJ1wm{utl'Ui~11lJ~ I'U II'l'j!'\l'Ui~11lJ~ t!1:I~fllllJ~'U l~mQ~tI~tltl1:l~ 46.35 6.74 t!1:I~77.30 l~tlJl11Unt!,r~ 1'l1lJri'liu '\llnf11'jI'l~!Wm!Ufll1~tI'l'U~'U I~ rOl'Ul'U 416 Itll C)f!1:l1t!'l1:I~1'l~!titln'\lln~n'I:JW~'Vl1~-ffW211'U1'VItJ1'Utl~!C)f1:l£fllllJLll:JJ1'j fI'1'Ums Ll~l~!tl'U IC)flfIU'j~!tlLl 11:I!ULlt!1:I~Ifl~!'ULl t!1:I~f11'j!'\l'1tlj'1'Um'l11'jm1:ll~!1'l1t1lJ'\llml"l~tlt!iI~ I~ rOl'Ul'U 8 ItllC)f!1:I1'l ~'lM'j,rLlfitl PCSMA53 CCSMA59 SSTBA15 SSTBA16 SSSMA5 SSTBA26 KBSMA65 !W~ KSSMA80 !rltl!1'l1t1lJ!~tl~I'l~!titln'l'Um'j1:l~mtlJl!f1titlU1:Itl~!~tl'lM l~m:JJ1W 108 !C)fml~tllJ1:I~Hll'l'j Q~vl'U~ltlm:JJ1I'l'j 2 lJ1:Iilill'l'j u'Uri'wll!l"l~tlt!iI~ 40 ~l ~!f11~ ~ . U 'Utltl~ IT'UftlU~11 ~~fl1:leu fl1:llJVI'Ul116 m'j N !C)f'U~!lJ I'l'j t!1:I~Q ~vl 'ULll'j 1:1~meu 'Ul'U 1:Itltl'Utl~ •~ . t!ufll1~tlm:JJ1I'l'j!'Vhtl'Uu'U!l"l~tlt!iI~ 40 ~ll1m~'\llt1tl~'l'U~'U ('If~~'U Ifl 'j 1'If) t!1:I~1~i'Utl1111'j '" ~ • . v mn 'lUlT'UftlU~11~~ fl1:ltlUfl1:llJVI'Ul150 m'jl~!C)f'U~!lJl'l'j 'I'11f11'j'VI~1:Itl~3 cih t!1:I~'l,*Lll'j1:l~mtl • 'J/ jI jI , 11l!f1ti ail 1:t1l~ !~tltl~l~!~ tllQ ~vl'U!l"l~ tit! iI~ ftl11 i' U'If~fllUfllJ n ru nu LlfIll~ 'VI~ 1:tl1 ~l1tlW 11n lJ q q q q qJ ~ ~ 25-28 tl~fl'1!C)f1:l!;tlLlfllllJ~'U-fflJl4''VI11~tltl1:l~ 70 !1J'U!d1:l15 1''U 'W1J';hJJf11'jl'llt1'Utl~!l"l~tlt!iI~'\lln n 1'j Q ~ vl 'U~ 1 tI u U fll1 !1 tI I e IC)f!1:I1'lPCSMA53 CCSMA59 SSTBA15 SSTBA16 SSSMA5 SSTBA26 KBSMA65 t!1:I~ KSSMA80 '\lln~tl1rYtlU'Utltl~lT'UftlU~11~~~tltl1:l~ 81.6770.038.33 II 35.8440.023.35 28.33 !W~ 18.38 19l1lJrl'1~1Jml~~fldh.!~h.!~fltJi:l~ 92.5090.8380.064.1757.50 'U o cv dd. I 1.cS QI 0 Q.I 3J tI 50.85 44.18 Ui:l~ 27.50 19l1lJi:l1\9l1J'YIlJfl11lJUI9lf119l1~fltJ1~lJ'l..W~nmu(P <0.05) '\'I1f1f1i:lfl~'\'Ii:l'YI'j'Hl'U u • U1J1Jfffl~f1'j 1\9l'Vl1J!C}fi:lrl'u 1Jfl-Yj~ tJ111U1'1...1 u 1f11J'1nw 'lffl~!lI \9l1J'1rnu ~1'1...1 1J'1...1'\Jfl~ff1U i1Ui:l ~ ff1U li'ltJrl'li1 Ui:l~'lffl~!lI \9l'\Jfl~lPifllJff~l~!~U 1'\J19l1lJtbrl'li 1 !rlflfff1lJ 1fflJ1J'11'Y11f~f1 'ji'YItJl'\Jfl~ U1Jfl-Yj~tJ~lU1U 8 'ln iC}fWI9lPCSMA53 CCSMA59 SSTBA15 SSTBA26 SSTBA16 KSSMA80 dodo 0 .:!I KBSMA65 Ui:l~ SSSMA5 'Vl1Jfl11lJ'\1i:l1f1'\1l:lltJ'\Jfl~fff1i:l'\Jfl~U1Jfl'YI!'jtJ '\'11'1...11'1...1 4 fff1i:l flfl • • . t 0 ~ c:/ .dt Pseudomonas Serratia Chromobactenum Ui:l~ Chryseobactenum '\'11n '1...1'1...1 '1...1 llJ 11! fl 'j 1~ '\1!'Vlfl '\11 rl'1~1J'W1fl1lifl1'Y1~'\Jfl~ 16S ribosomal RNA gene Ui:l~!rlmllitJ1J!-YjtJ1Jfl11lJmijflunmrmqji:l'\'l1f1 9J .cS ~ QI i 1'1...1 '\Jfllq! i:l GenBank lJ fl11lJ !'\1lJ fl '1...1 f11J Pseudomonas aeruginosa KVD14-MG Serratia marcescens 7/18r Chromobacterium pseudoviolacium LMG3953 Chryseobacterium gleum ALR- 8 Chromobacterium piscinae LMG3947 Pseudomonas jluorescens DKPl Chryseobacterium iJ indologenes MUT2 Ui:l~ Chromobacterium piscinae LMG3947 'jfltJi:l~ 98.096.899.399.798.3 99.8 99.2 Ui:l~ 96.6 19l1lJrl'1~1J ~I dI ufll'jfl'mn 2558 PATCHARIDA KHUMPUMUANG : BACTERIA IN SOIL CAPABLE OF DESTROYING STRUCTURE OF CASSA VA MEALY BUGS. THESIS ADVISOR: ASST. PROF. SUREELAK RODTONG, Ph.D. 155 PP. CASSA VA MEAL YBUGS/SOIL BACTERIAl STRUCTURE OF MEALY BUGS Mealy bug is important pest insect of cassava, could destroy plant by suck to eat the nutrient from every the part of the plant were depressed plant dies. The control cassava mealy bugs by soil bacteria have tendency economical and sustainable methods. From the study of structures and chemical components of cassava mealy bugs that could serve as nutrients of bacteria and the possibility of using selected isolates of soil bacteria for destroying structures of cassava mealy bug, that destroy the plant in Northeastern Thailand. Found species, pink mealy bug (Phenacoccus manihoti Matile-Ferrero) higher than three species, green mealy bug (Phenacoccus maderiensis Green), jack beard mealy bug (Pseudococcus jackbeardsleyi Gimpel & Miller), and striped mealy bug (Ferrisia virgata Cockerel). Pink mealy bugs were studied, have cuticle layer size, 3-4 urn. Found the structure of open pores can destroy by using soil bacteria such as anterior and posterior pores (ostioles), several pores for producing wax filament, abdomen pores (circulus) and respiration pores (spiracles) have diameter 2-59,2-7, 17-20, and 9-30 urn, respectively. Chemical compositions of the body of mealy bug found total carbon, total nitrogen, and moisture content 46.35, 6.74, and 77.30% by dry basis, respectively. A total of 416 bacterial isolates from soil samples were collected and screened by morphological characteristic, producing protease lipase and chitinase enzymes, and can grow in cassava mealy bug broth. Eight isolates ofPCSMA53, CCSMA59, SSTBA15, SSTBA16, SSSMA5, SSTBA26, IV KBSMA65, and KSSMA80, after sprayed 2 ml of bacterial suspension (108 cells/ml) with 0.85% normal saline on third instar stages of forty cassava mealy bug feed on peak of cassava stem, the area 16 square centimeter and compared with in soil, the area 50 square centimeter, 70 humidity at 25-28 °C for 5 days (triplicate test).
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