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Isozymic Studies on Selected Species of Pteris from India

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Isozymic Studies on Selected Species of Pteris from India id2612421 pdfMachine by Broadgun Software - a great PDF writer! - a great PDF creator! - http://www.pdfmachine.com http://www.broadgun.com BBAprili i20oo10 ISSN : 0974 - 7427 Volume 4 Issue 1 CCHHEEMMAInI ISSndTTian RRJourYYnal Trade Science Inc. Regular Paper BCAIJ, 4(1), 2010 [54-57] Isozymic studies on selected species of Pteris from India M.Johnson2*, V.Irudaya Raj2, S.D.Rajkumar1 ’ 1Department of Botany, St. Andrew s College, Gorakhpur - 273 001, UP, (INDIA) ’ 2Department of Plant Biology and Plant Biotechnology, St. Xavier s College (Autonomous), Palayamkottai, Tamil Nadu, (INDIA) E-mail : [email protected] Received: 10th December, 2009 ; Accepted: 20th December, 2009 ABSTRACT KEYWORDS The present study was aimed to illustrate the isoperoxidase banding profile Isozymes; for some species of Pteris from India. The standard technique of vertical Iso-peroxidases; poly acrylamide gel electrophoresis was employed to demonstrate the band- Pteis argyraea; ing profile and the gels were stained for isoperoxidase enzyme system. In Pteris otaria; this enzyme system, a total of thirty eight bands were scored in thirty one Pteris vitata; different positions with eight active zones. The present study revealed the Pteris biaurita; biochemical positions of the Pteris complex. The similarity index and varia- Pteris confuse. tions between these five species are reported with reference to isoperoxidase profiles. 2010 Trade Science Inc. - INDIA INTRODUCTION play an important role in phytoremediation throughout the world. The Chinese Brake fern Pteris vittata have Ferns and fern allies, collectively called Pterido- been studied extensively for its potentiality in phytes, are the first successful group of land plants with- hyperaccumulation of Arsenic[2]. In a recent study the out flowers and seeds. India is one of the important spores of related species Pteris confusa T. G. Walker megabiodiversity countries with the presence of about and P. argyraea T. Moore shows reverse response in 1200 species of Pteridophytes which are growing on heavy metal tolerance during germination[3]. There are the high altitude mountains like the Western Ghats and 15 species of Pteris in south India[4]. General taxo- Himalayas. Pteris is a highly evolved tropical fern ge- nomical, cytological and phytochemical studies on south nus with the presence of several species complexes like Indian Pteris have been done[4-6] and the results show Pteris quadriaurita, P. cretica and P. vittata with the high degree of morphological, cytological and chemi- several morphotypes and cytotypes. Walker[1] has ob- cal diversity of south Indian species of Pteris. Isozyme served introgression resulting in the formation of hybrid analysis reveals the systematic positions of several taxa swarm in nature. Majority of the species are sexual dip- with inexpensive technique[7,8]. In 1960s, biochemical loids or tetraploids and some species are diploid or methods based on seed, leaves protein or enzyme elec- triploid apogamous. With the chance for easy hybrid- trophoresis were introduced, which proved particularly ization, several morphotypes and cytotpyes occur in useful in analysis of genetic diversity as they reveal differ- nature. Ferns in general, Pteris species in particular, ences between seed storage protein encoded by different BCAIJ, 4(1) April 2010 M.Johnson et al. 55 Regular Paper alleles at one (allozymes) or more gene loci (isozymes). the selected ferns in this enzyme system. Zone 3 con- Use of biochemical methods eliminates the environmental tained two bands in two different positions. Band PRX influence. These methods are being used as complemen- 31 (0.275) showed its presence in P. otaria and P. tary strategies to traditional approaches for assessment of confusa. PRX 32 (0.284) was shared by P. argyraea genetic diversity. The analysis can be performed at any and P. biaurita collected from Kothayar (2 x). Zone 4 growth stage using any plant part and it requires only small showed three bands in three different positions. PRX amounts of materials. Following the advances in molecular 41 (0.304) showed its unique presence in P. otaria only. biology in the last decade a variety of different methods PRX 42 (0.375) was unique to P.argyraea and PRX have been developed for analysis of genetic diversity. In 43 (0.400) was restricted with diploid P. biaurita col- order to know the genetical diversity among the spe- lected from Kothayar. Thus the zone 4 played a key cies of Pteris from South India, isozyme analysis has role in characterization of P. otaria, P. argyraea and been done in some common species of Pteris particu- P. biaurita. Zone 5 contained three bands in three dif- larly from Pteris quadriaurita complex along with an- ferent positions. PRX 51 (0.422) was unique to triploid other unrelated species Pteris vittata. apogamous P. biaurita collected from Kodaikanal botanic garden. PRX 52 (0.481) was restricted to P. MATERIALS AND METHODS otaria and PRX 53 (0.491) for P. argyraea. Thus the zone 5 played a key role in the characterization of trip- Pteis argyraea, Pteris otaria, Pteris vittata, loid apogamous P. biaurita collected from Kodaikanal Pteris biaurita and Pteris confusa were collected from botanic garden. Zone 6 showed six bands with six dif- different localities of the Western Ghats, South India ferent position of expression. PRX 61 (0.534) was and established in the green house at Centre for shared by P. biaurita (Kothayar populations) and P. ’ Biodiversity and Biotechnololgy, St. Xavier s College, argyraea. PRX 62 (0.555) was commonly present in P. Palayamkottai, Tamil Nadu, India. The croziers were otaria and P. biaurita (Kothayar). PRX 63 (0.567) harvested from mother plants and used as the experi- was restricted to P.confusa, PRX 64 (0.574) was unique mental materials for the enzyme isolation. 500 to 1000 to P. argyraea, PRX 65 (0.579) was present only in P. mg of freshly harvested young croziers were taken and vittata and PRX 66 (0.588) was exclusive to P. biaurita homogenized with 3.5 ml of ice-cold homogenizing buffer (Kothayar). Zone 7 also expressed six bands in six dif- (0.1M phosphate buffer (pH 7.0)) and centrifuged at ferent banding patterns. PRX 71 & 6 (0.601 and 0.683) 12,000 rpm for 10 min. The supernatant was subjected was unique to P.viattata, PRX 73 (0.629) was restricted to electrophoresis (PAGE) as described by Anbalagan[9]. to P. argyraea, PRX 74 (0.634) showed its presence in For the detection of isozymes on the gels, the staining P. biaurita (Kodaikanal botanic garden populations), solution, were prepared as per Smila et al., method[10]. PRX 72 (0.613) was shared by P. confusa and P. After the electrophoresis, the gels were incubated in vittata. Zone 8 recorded six bands with different band- the staining solution for few minutes under dark condi- ing positions. PRX 81 and 3 (0.710 and 0.748) were unique tion till the clear bands appeared. The gels were fixed to P. vittata. PRX 84 and 6 (0.760 and 0.779) were re- with 7% acetic acid solution for 30 min, washed with stricted with P. argyraea. PRX 85 (0.768) to P. biaurita distilled water and banding patterns were documented Kodaikanal botanic garden population. PRX 82 (0.723) using the Vilber Loubermat gel documentation system showed the common presence in P. argyraea and P. (Germany) and the similarity index and variations were vittata. Zone 9 showed four bands in four different calculated using the Biogene software (Germany). positions in the enzyme system. PRX 91 and 4 (0.809 and 0.890) were unique to P. biaurita from Kodaikannal RESULTS AND DISCUSSION Botanic Garden. PRX 92 (0.818) was restricted to P. confusa and PRX 93 (0.845) was shared by P. biaurita Isoperoxidase enzyme system showed the multiple (KBG) and P. argyraea. Zone 10 showed a single band zones[8] of activity with different banding profiles. Zone PRX 101 (0.926) and it is restricted to P. biaurita from one and two failed to express the banding profile for KBG. Bio CHEAMn InIdSianT JoRurnYal 56 Isozymic studies on selected species of Pteris from India BCAIJ, 4(1) April 2010 Regular Paper TABLE 1 : Isoperoxidase banding pattern of some species of Pteris s a a a a a s e t t ) d e a s a e i i i i l t ) i a i u r r u r a x r a f f l G t u u a y h o t o n a t t i a a B r g r i i o v o o v r e K p c b b . f a ( K p . ( e P . P o R P P P s P I 0.275 PRX31 + + 0.284 PRX32 + + 1 0.304 PRX4 + 2 0.375 PRX4 + 3 0.400 PRX4 + 1 0.422 PRX5 + 2 0.481 PRX5 + 3 0.491 PRX5 + 1 0.534 PRX6 + + 2 Figure 1 : Isoperoxidase profiles of Pteris sps. P.o-Pteris otaria, 0.555 PRX6 + + P.c-Pteris confuse, P.b(KBG)-Pteris biaurita from Kodaikanal 0.567 PRX63 + Botanic Garden, P.b(Kot)-Pteris biaurita from Kothayar, 0.574 PRX64 + Tirunelveli, P.a-Pteris argyraea, P.v-Pteris vittata 5 0.579 PRX6 + bers of the P. quadriaurita complex and this morphol- 6 0.588 PRX6 + ogy based classification is supported by the present 1 0.601 PRX7 + isozymic profile analysis. In general simplified lamina is 2 0.613 PRX7 + + more advanced one when compared with the more pin- 3 0.629 PRX7 + natifid fronds. Population of P. biaurita L. from 4 0.634 PRX7 + Kodaikanal Botanical Garden is with high genetic vari- 5 0.650 PRX7 + ability with the presence of eight bands when compared 6 0.683 PRX7 + with the population of the same species from Kothayar 1 0.710 PRX8 + which is with only four unmatched bands.
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