Visualization and Interpretation of Plant Isozymes

Wendel, Jonathan F.; Weeden, N. F.

doi:10.1007/978-94-009-1840-5_2

Cited by 947 publications

(651 citation statements)

References 130 publications

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“…Electrophoresis was carried out using 14% hydrolysed potato starch gel in 'Genei' horizontal migration chamber at constant current of 24 mA for first 30 min followed with 34 mA. The gels were stained following the method of Veech (1969) for peroxidase and Wendel and Weeden (1989) for other enzymes.…”

Section: Methodsmentioning

confidence: 99%

In vitro Callusing and Regeneration in Trifolium resupinatum-A Fodder Legume

et al. 2006

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SummaryIn vitro regeneration protocol was developed in Trifolium resupinatum, an important fodder legume in Northwestern India. Out of 6 explants (leaf, petiole, hypocotyl, cotyledon, collar and root) tried for callus induction in 2 different media, good response was observed from hypocotyl and root explants in 'A' medium. Successful regeneration was obtained from callus induced from hypocotyl, cotyledon and root in 'A' followed with subculture in 'E' medium. One of the regenerated plantlet was characterized and compared with mother plant for isozyme-banding pattern. Wide variation for isozyme banding pattern for SOD, Peroxidase and GOT enzyme indicates the regenerant to be a somaclonal variant. Thus, in vitro regeneration will help in widening the genetic base of the species.Key words Trifolium resupinatum, Fodder, Regeneration, Tissue culture.Plant tissue, cell and protoplast cultures are useful tools for crop improvement, especially as supplementary means of inducing variability through tissue culture instability and somaclonal variations (Scowcroft 1984). Trifolium resupinatum (Shaftal or Persian clover) is an important cultivated winter annual fodder legume grown widely in subtropical and sub temperate areas of India. The species has previously been frequently introduced in different parts of Europe and cultivated as fodder crop in northwestern subtropical zones. It is a multicut crop and is favoured by farmers for its high quality, good yield and tolerance to biotic and abiotic stress.Narrow genetic base in this crop hampers its genetic improvement programme. The conventional breeding efforts have failed to generate variability for making effective selection. In this study, efforts were made to develop and characterize somaclonal variants in this crop by using in vitro tissue culture methods. Materials and methodsHealthy seeds of T. resupinatum (SH 97-49) were surface sterilized using 0.1% HgCl 2 for 2 min followed by 4-5 rinsing with sterile water and germinated on basal MS medium (Murashige and Skoog 1962) containing 3% sucrose and 0.7% agar without any growth regulators. Six explants viz., leaf, petiole, hypocotyl, cotyledon, collar and root were taken from 21-23 d old seedlings. Two combinations of auxin and cytokinin in the medium were studied for callus induction efficiency. The medium 'A' contained L2 basal medium (Phillips and Collins 1984) supplemented with 0.05 mg/l NAA and 0.1 mg/l BAP, whereas medium 'A-1' contained L2 basal medium supplemented with 0.1 mg/l BAP. Initially the explants were kept in dark for callus induction and after 5 d of inoculation the cultures were provided with a photoperiod of 10 h. After shoot emergence, the light

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Section: Methodsmentioning

confidence: 99%

In vitro Callusing and Regeneration in Trifolium resupinatum-A Fodder Legume

et al. 2006

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“…Crude extract was made by grinding each sample with 5 ml of 75 mM Tris-HCl buffer, pH 7.5 containing 5 % glycerol (w/v), 5 % PVP-40 (w/v), 14 mM mercaptoethanol (0.1 % v/v), 50 mM Na-salt, 10 mM dithiothreitol (DTT) and 0.1 % bovine serum albumin (w/v) (Wendel and Weeden 1989). The homogenate was centrifuged at 10, 000 g for 20 min at 4°C and the supernatant was used for identification of different isoforms of SOD.…”

Section: Sod Enzyme Extractionmentioning

confidence: 99%

Analysis of biochemical variations and microRNA expression in wild (Ipomoea campanulata) and cultivated (Jacquemontia pentantha) species exposed to in vivo water stress

Ghorecha

Patel

Ingle

et al. 2013

Physiol Mol Biol Plants

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The current study analyses few important biochemical parameters and microRNA expression in two closely related species (wild but tolerant Ipomoea campanulata L. and cultivated but sensitive Jacquemontia pentantha Jacq.G.Don) exposed to water deficit conditions naturally occurring in the field. Under soil water deficit, both the species showed reduction in their leaf area and SLA as compared to well-watered condition. A greater decrease in chlorophyll was noticed in J. pentantha (~50 %) as compared to I. campanulata (20 %) under stress. By contrast, anthocyanin and MDA accumulation was greater in J. pentantha as compared to I. campanulata . Multiple isoforms of superoxide dismutases (SODs) with differing activities were observed under stress in these two plant species. CuZnSOD isoforms showed comparatively higher induction (~10-40 %) in I. campanulata than J. pentantha . MicroRNAs, miR398, miR319, miR395 miR172, and miR408 showed opposing expression under water deficit in these two plant species. Expression of miR156, miR168, miR171, miR172, miR393, miR319, miR396, miR397 and miR408 from either I. campanulata or J. pentantha or both demonstrated opposite pattern of expression to that of drought stressed Arabidopsis. The better tolerance of the wild species (I. campanulata) to water deficit could be attributed to lesser variations in chlorophyll and anthocyanin levels; and relatively higher levels of SODs than J. pentantha. miRNA expression was different in I. campanulata than J. pentantha.

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“…For gels staining, protocols of Scandalios (1964) were used for α and β-Est. ; Wendel & Weeden (1989) for both Ao and Acp; Weeden & Wendel (1990) for Adh; Jonathan & Wendell (1990) for Mdh & Heldt (1997) for Px. After run finished, gels were washed two or three times with tap water; fixed in ethanol: 20% glacial acetic acid (9:11 v/v) for 24 hr and photographed.…”

Section: Methodsmentioning

confidence: 99%

Preliminary Authentication of Some Acacia L. Species in Taif Highlands

2015

Egypt. J. Bot.

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LTHOUGH their wide distribution in arid lands and their many uses that include fodder and fuel besides the environmental values of soil stabilization, Acacia species are vulnerable to elimination in Saudi Arabia. In this study, seed morphology and patterns of their coat surface sculpture as revealed by scanning electron microscopy besides both seed proteins and seven isozymes profiles were employed for the discrimination and authentication the vulnerable Saudi Arabian Acacia collected from the western region of the kingdom. The scanning electron microscopic study displayed diversity in shape, dimensions, color, central aerole features and coat topography of seeds among different species to be characteristic for each species. Seed protein and isozyme profiles showed high variability among studied species. The UPGMA phenogram and genetic similarity analysis based on combination of seed morphology, protein and isozyme patterns confirmed the extensive genetic diversity existed in Acacia species.

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Visualization and Interpretation of Plant Isozymes

Cited by 947 publications

References 130 publications

In vitro Callusing and Regeneration in Trifolium resupinatum-A Fodder Legume

In vitro Callusing and Regeneration in Trifolium resupinatum-A Fodder Legume

Analysis of biochemical variations and microRNA expression in wild (Ipomoea campanulata) and cultivated (Jacquemontia pentantha) species exposed to in vivo water stress

Preliminary Authentication of Some Acacia L. Species in Taif Highlands

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