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Limited germplasm and narrow genetic base in the past has hinders wider applicability of Stylosanthes in India. However, build up in Stylosanthes germplasm in recent years provided an opportunity to evaluate and use them under different agro-ecological situations preferably targeting drought tolerance. Rate reducing resistance (RRR) allo-tetraploid lines of S. scabra and diploid S. seabrana, a newly introduced species, observed well suited in hard and cracking soils under complete rainfed condition. Genotypes of S. scabra were more tolerant to drought over lines of other species as evidenced by high leaf thickness, more proline accumulation, contents of malondialdehyde (MDA), sugars, starch and chlorophyll and low carbon isotope discrimination (CID) values. Both in control and stress conditions, a positive relationship (r = 0.465 and 0.328) was observed between specific leaf area (SLA) and CID. Earlier reports have emphasized measurement of CID as an indirect way of measuring the transpiration efficiency (TE) in Stylosanthes (Thumma et al. Aust J Agr Res 49:1039–1045, 1998). The negative relationship observed between TE and CID suggested low CID bearing lines would have high TE, useful traits to select lines growing better in dry environments. Thus, selected lines having low CID provided better scope for the introduction of this important range legume for semi- arid regions of India.
Many species of the genus Cenchrus, with different ploidy are important components of world grasslands. In the present investigation, influence of ploidy on leaf water potential (Ψleaf) and photosynthetic characteristics under water-stress condition was studied in 2-month-old plants of eight Cenchrus species representing three ploidy and two life spans. Stress was imposed for 2, 4, 6, and 8 days by withholding watering. Decrease in water potential, chlorophyll content, photosynthetic rate (PN), stomatal conductance for CO₂ (gs), and carboxylation efficiency (PN/Ci) occurred in all species. Nevertheless, perennial and diploid C. setigerus maintained the highest PN/Ci at maximum stress indicating its higher assimilation potential. Photosynthetic water use efficiency (PN/E) in general decreased with increase of water stress; however, at low stress, reverse trend was observed in diploid C. prieurri, C. setigerus, and tetraploid C. glaucus. Results indicate that relative reduction in photosynthetic characteristics was more in annual diploid than perennial diploid and tetraploid. Stress tolerance in terms of plant height, rolling and wilting of leaves also indicated better adaptability of tetra and hexaploid species over diploid. Moreover, recovery of different physiological traits was more pronounced in perennial tetraploid and hexaploid over annual diploid, thus making them suited more to drought conditions. Analysis of variance results showed direct correlation (r = 0.675) of ploidy with gas exchange parameters indicating better impact of this on physiological performance of different species of Cenchrus under water stress. This is the first report where influence of ploidy levels on photosynthetic characteristics and overall responses to moisture stress is studied in a tropical grass species.
Three families of proteinase inhibitors, namely, serine, cysteine (thiol) and aspartic (carboxyl) were examined for their inhibitory effects on growth and development of Indian alfalfa weevil (Coleoptera: Curculionidae). Proteinase inhibitors are considered as a part of alternate strategy to control the herbivorous insect as they inhibit the digestive enzymes of the insects. Larval leaf feeding, survival, pupation and adult emergence were significantly decreased by pHMB, (p-hydroxy-mercuribenzoic acid), cystatin and E-64 (trans-epoxysuccinyl-L-leucylamido-( 4guanidino)-butane) belonging to cysteine class of proteinases, at a concentration of 0.1 and 0.5%. Serine and aspartic classes of inhibitors have low detrimental effects on larvae. The results demonstrate the inhibitory response of specific proteinase inhibitors on alfalfa weevil larval leaf feeding, survival, pupation and adult emergence. Weevil resistant species, namely, Medicago scutellata showed high level of leaf consumption under forced feeding in vivo bioassay indicated the presence of resistance factors other than proteinase inhibitors.
Cenchrus (family Poaceae) is an important component of major grass covers of the world. Largely it is apomictic and both annual and perennial species exist in nature. Variations in contents of malondialdehyde, proline, specific leaf area and carbon isotope discrimination for drought tolerance were estimated among eight prominent species of Cenchrus. Simultaneously, genetic variations were also estimated by employing 187 RAPD primers. Of these, 23 primers did not react, 2 performed poorly and 7 produced many non-scorable bands and one primer yielded a single monomorphic band. Rest of the 154 primers generated one or more unambiguously scorable fragments. Twelve hundred and four of the 1,296 putative loci were polymorphic (93%) between at least one pair-wise comparisons among eight species. Dice coefficient and neighbor-joining algorithm analyses showed clustering patterns that fit with the known habitat of the species except perennial, C. myosuroides which formed a node between two annuals species. When these species were subjected to water stress tolerance test, a correlation (r = 0.612) between specific leaf area (SLA) and carbon isotope discrimination (CID) and difference in levels of drought tolerance based parameters among eight species were observed. Of the eight species investigated two annuals viz., C. biflorus and C. echinatus showed highest level of genetic similarity which was also evident from the similar levels of SLA, MDA, proline contents and carbon isotope discrimination values observed in these two species.
In this paper, we describe a study concerning the determination of phenylalanine ammonia lyase (PAL) activities leading to decline in disease progress caused by Rhizoctonia solani after application of salicylic acid (SA). Two applications of 1.4 mM SA (pH 6.5) followed by inoculation with Rhizoctonia solani resulted in a quantitative change in polyphenol oxidase (PPO), peroxidase (PRX) isoforms and increase in PAL activities from 4.38 to 19.48 unit g⁻¹ (FM) h⁻¹ in Bundel-1, UPC-4200 and IFC-902 cowpea genotypes. Increase in PAL activities was further observed specifically in UPC-4200 when plants were exposed with Rhizoctonia solani spores. Total soluble proteins did not change after SA treatment, however they were significantly increased in SA sprayed-inoculated UPC-4200 genotype. Of the ten detected isoforms of polyphenol oxidase, isoforms 7 and 10, and isoform 4 of peroxidase showed increased activities by SA application. The disease symptoms measured as areas under progress curve (AUDPC) indicated less A value in SA sprayed Bundel-1 and UPC-4200 genotypes over their controls.
Among the many Stylosanthes species, Stylosanthes scabra, a range fodder legume, performs better under limited water condition. In the present investigation, thirty-four accessions of S. scabra were assessed under limited water condition, for various morpho-physiological characters associated with drought. In general, S. scabra exhibited better tolerance to drought, as evidenced by high leaf thickness and greater accumulation of proline, and malondialdehyde (MDA) in water stress condition. Transpiration efficiency (TE) was high, in both control and water stress conditions and positively correlated with root, shoot, and total dry matters, in both control and stress conditions (r 2 = ranged from 0.589 to 0.961 in control and from 0.351 to 0.985 in stress). Of these, 25 accessions were assessed for estimation of genetic diversity, employing random amplified polymorphic DNA (RAPD) markers. A total of 210 RAPD bands, obtained with 32 primers, revealed high polymorphic information content (0.49) and marker index (4.41). Dendrogram analysis indicated close proximity among the accessions of S. scabra. These accessions were clustered in high similarity range (84.01–98.36 %). Accession IG-366A separated from other clusters at 85.62 % similarity level. RAPD marker system revealed 13 accessions exhibiting >90 % genetic similarity while the other accessions exhibited similarity ranging from 68 to 90 %. A higher level of genetic similarity which was also evident from the similar levels of TE, biomass production, root/shoot ratio, MDA, proline contents and drought tolerance index, indicated a cause–effect relationship among them. Results also indicated that among the accessions, S. scabra rate-reducing resistance allo-tetraploid lines were better suited for hard and cracking soils, under complete rain-fed condition.
The present study was aimed at improving sprouting and establishment of bud chip seed stocks of sugarcane cultivar CoSe 92423 by pre-planting soaking in growth-promoting chemicals viz ethephon (0.1 g dm⁻³) and calcium chloride (1 g dm⁻³) along with water-soaked control for 24 h. Treated bud chips recorded higher bud sprouting, shoot height, root number, fresh weight of leaves, shoot and roots, and plant vigor index. In both the treatments, reducing sugars contents, acid invertase, and ATPase activity increased in developing sprouts; increase was about 86.5 and 40.7% in reducing sugars, 28 and 70% in acid invertase, and 15 and 23% in ATPase activities over control by ethephon and calcium chloride treatment, respectively. Reducing sugars contents and activity of acid invertase and ATPase enzymes of sprouted buds exhibited significant positive correlation with bud sprouting and plant vigor index. These findings indicate that soaking of bud chips in growth-promoting chemicals viz ethephon (0.1 g dm⁻³) and calcium chloride (1 g dm⁻³) solutions helps in enhancing bud sprouting, root growth, and plant vigor by altering some of the key biochemical attributes essential for the early growth and better establishment of bud chips under field conditions which is otherwise poor in untreated chips.
The present study employed a sand culture experiment with three levels of zinc viz., 0.065 (control), 65.0 and 130 mg l⁻¹ Zn (excess) as zinc sulfate, respectively, in sugarcane (Saccharum spp.), cultivar CoLk 8102. The results indicated growth depression, dark green leaves, decreased root number and length and sharp depression in mitotic activity of roots due to high doses of Zn (65 and 130 mg l⁻¹); effects were significant at 130 mg l⁻¹ Zn supply. The endogenous ion contents measurements revealed roots to be the major sink for excess Zn with lower amounts in leaves of sugarcane plants. High level of Zn decreased total phosphorus in leaves and increased it in roots. Fe and Cu content decreased, while, Mn increased in sugarcane plants due to high Zn in the growing medium. Plants experienced oxidative stress when exposed to higher levels of zinc. Biochemical investigations indicated high level of hydrogen peroxide, malondialdehyde contents with high chlorophyll a, b and carotenoids contents and activity of superoxide dismutase, catalase and peroxidase enzymes under high Zn conditions. These findings confirm suggest that excess Zn adversely affects root growth and mitotic efficiency, enhances chromosomal aberrations and increases growth and nutrient accumulation abnormalities, as well as oxidative stress.
Aconitum violaceum Jacq. is an important medicinal species used for various health ailments including renal pain, rheumatism and high fever. In the present report, a reproducible in vitro regeneration system for Aconitum violaceum Jacq. has developed from the nodal segment of the plant. Induction of shoot buds was achieved on basal Murashige and Skoog (MS) medium. The shoots were elongated on MS medium supplemented with 0.5 μM 6-benzylaminopurine (BAP) and 0.1 μM α-napthaleneacetic acid (NAA) and subsequently transferred to rooting medium. In vitro grown microshoots of A. violaceum were encapsulated in the alginate beads. The success rate of their re-growth was found to be approximately 85.43 %. Of the encapsulated microshoots, 39.86 % exhibited formation of multiple shoots following re-growth on plant growth regulator free MS medium. Healthy root formation was observed in all microshoots following 2 weeks of transfer on half-strength MS medium containing 0.1 lM indole-3- acetic acid (IAA) and 1.0 μM α-naphthalene acetic acid (NAA). These plants were subsequently transferred to pots containing a mixture of soil, sand and compost (1:1:1 v/v), and same were then shifted in the greenhouse with 87 % survival rate. The molecular analysis was carried out using 35 random amplified polymorphic DNAs (RAPD) primers and 25 inter simple sequence repeats (ISSR) primers. Cluster analysis of the RAPD and ISSR profile revealed an average similarity coefficient of 0.966 and 0.974, respectively, confirming genetic stability of tissue culture-raised (TR) plants and synthetic seed-derived plants (SR). The phytochemical analysis of tissue culture-raised and synthetic seeds-derived plants showed higher aconitine content than control plant. The propagation protocol developed in this study provides a basis for germplasm conservation and harnessing the medicinally active compounds of A. violaceum.
Genetic stability and phytochemical analysis of in vitro established plants of Picrorhiza kurroa Royle ex Benth, have been carried out. Random amplified polymorphic DNA (RAPD) and inter simple sequence repeats (ISSR) markers were used to assess the genetic fidelity of tissue culture products including three adventitious shoots from three calli and 6 months old tissue culture raised plants growing in green house condition with mother plant. Apparent genetic variation was detected in the five types of plant materials. The percentage of polymorphic bands in the RAPD and ISSR analysis were 16.25 and 14.54 %, respectively. The genetic similarity was calculated on the basis of RAPD and ISSR data among the five types of plant materials and were ranged from 0.5 to 1.0 (mean 0.75) and 0.47 to 1.0 (mean 0.73), respectively. The similarity coefficient by both RAPD and ISSR analysis revealed that differences between tissue culture raised plants and mother plant was not remarkable, but notable differences were observed among three adventitious shoots regenerated from three calli. The phytochemical analysis of tissue culture raised products showed higher secondary metabolite (picrotin and picrotoxinin) content as compare to mother plant. The information gained on genetic stability/variability will be valuable for the large scale propagation and secondary metabolite production of P. kurroa.
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