Aenhenrya rotundifolia is a critically endangered terrestrial jewel orchid. It is monotypic and endemic to evergreen forests of southern western ghats of India. In the present study, identification of this plant species is validated with DNA barcoding using matK and rbcL chloroplast markers. Further, germ-free juvenile axillary bud explants were cultured on Mitra medium supplemented with different kinds of cytokinins like 6-benzyladenine, 6-furfurylaminopurine, N 6 -(D 2 -isopentyl) adenine, thidiazuron, zeatin and meta-topolin as well as auxins such as a-naphthaleneacetic acid, indole-3-acetic acid and indole-3-butyric acid at different concentrations and combinations for successful proliferation and establishment in vitro. After 12 weeks of culture, axillary bud explants produced an average of 30.12 ± 0.71 shoots per explant, 3.87 ± 0.06 cm shoot length, 1671 ± 2.82 mg fresh mass of proliferated shoots with a proliferation frequency of 100% on Mitra medium supplemented with 6.20 lM metatopolin and 2.25 lM thidiazuron. No root formation was observed in in vitro proliferated microshoots. However, tiny hair like projections were observed in some elongated shoots on Mitra medium pertaining to 5.37 lM NAA. The tiny hair like structure bearing plantlets were hardened and acclimatized with 100% survival rate in the polytunnel chamber. After 8-10 months of establishment ex vitro, flowering was observed. Additionally, the genetic fidelity of in vitro derived plants was tested with ISSR and SCoT marker profiling. The test results revealed that the plants derived from the protocol has 99% genetic similarity to that of the donor mother plant. This study can be applied in forensic interventions of this species, describes the maintenance of germplasm in vitro and establishment of new viable population in its original habitats by restoring existing sites of this critically endangered jewel orchid.
Stem rust of wheat caused by Puccinia graminis Pers. f. sp. tritici Eriks. & E. Henn., is the most destructive disease of wheat worldwide as well as in Egypt. It can causes high sever losses of wheat crops over wide areas during epidemic years. Hence, this study was carried out to detect the virulence dynamic and diversity of P. graminis f. sp. tritici in different locations and studying the efficacy of stem rust resistant genes in Egypt seedling stage. Stem rust collections were obtained from infected wheat stems throughout the survey of wheat fields and nurseries in three locations (Sids, El Sharkia and El Nubaria) during 2011/2012 growin g season. Whereas during 2012/2013 growing season the samples collected from six locations (Giza, Sids, Tag El Aiz, Sakha, El Sharkia and El Nubaria). Based on race analysis of stem rust populations, and race determination by inoculating stem rust differential hosts, the phenotypic characterization of P. graminis f. sp. tritici during 2011/2012 growing season resulted in identification of 86 races from 22 successful samples, all of them showed 1.16% frequency. Race BBBBC was a virulent on all the tested Sr genes, except Sr MCN, whereas race TTTTK was virulent on all the tested Sr except Sr 24. On the other hand, analysis during the next growing season revealed that, 123 races with a frequency ranged from 0.81% to 2.43% were identified. Race groups BB-, LG-, BJ-and TT-were common at the tested locations during the two growing seasons. Regarding stem rust resistant gene efficacy during the study Sr24, Sr38 and Sr31 exhibit the highest efficacy% (95.34, 91.86 and 87.21 respectively) during the two seasons. Thus, deployment of effective Sr genes such as Sr24, Sr38 and Sr31 in single cultivar through gene pyramiding has paramount importance as the additive effects of several genes gives the cultivar a wider base stem rust resistance along with periodic race survey.
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