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1.
The effects of incorporating 5-aminolevulenic acid (ALA) into the priming solution on low-temperature germination and emergence percentage performance of red pepper (Capsicum annuum cv. Sena) seeds before and after seed storage were investigated. Seeds were primed in 3% KNO3 solution for 6 days at 25 °C in darkness containing 0 ppm, 1 ppm, 10 ppm, 25 ppm, 50 ppm or 100 ppm ALA. Following priming, seeds were either immediately subjected to germination and emergence tests at 15 °C or stored at 4 °C or 25 °C for 1 month after which they were subjected to germination and emergence tests at 15 °C. Priming pepper seeds in the presence of ALA improved final germination percentage (FGP) and germination rate (MGT) at 15 °C compared to non-primed seeds. The highest FGP was obtained from seeds primed in the presence of 25 ppm and higher ALA concentrations while the highest MGT was obtained from seeds primed in KNO3 supplemented with 10 ppm ALA. Emergence percentages were the highest for the seeds primed in the presence of 25 ppm ALA and 50 ppm ALA while non-primed seeds had the lowest emergence percentage. Highest emergence rates (MET) and heaviest seedlings were also obtained from seeds primed in KNO3 supplemented with 50 ppm ALA. Although all priming treatments improved germination and emergence performance of pepper seeds at 15 °C following 1 month of storage under two different temperatures, inclusion of 25 ppm and 50 ppm ALA into the priming solution resulted in higher germination and emergence percentages and faster germination and emergence compared to seeds primed in KNO3 only and non-primed seeds. These results indicate that priming seeds in 25 ppm and 50 ppm ALA incorporated into the KNO3 solution can be used as an effective method to improve low-temperature performance of red pepper seeds and that these seeds can be stored for 1 month at 4 °C or 25 °C and still exhibit improved germination and emergence performance at 15 °C.  相似文献   

2.
This study evaluated the effects of priming on emergence responses of purple coneflower (Echinacea purpurea L. Moench) seeds. The seeds that differ in seed size were either primed with moistened vermiculite (solid matrix priming) or primed in non-aerated −0.5 MPa polyethylene glycol 6000 solution at 25 °C for 6 days (osmopriming), followed by air-drying to their initial moisture level. The tetrazolium staining tests indicated that both large and small seeds were biochemically viable. No notable difference in germination percentage was found between large and small seeds. However, extensive cavity was visible in portions of small seeds in comparison with large seeds. Large seeds accumulated more antioxidants and had greater activities of anti-oxidative enzymes than small seeds. They also had greater isocitrate lyase and malate synthase activities than small seeds. As a result, large seeds had higher emergence percentage and faster emergence speed as compared to that of small seeds. Both solid matrix priming and osmopriming increased emergence percentage and shortened mean emergence time of purple coneflower seeds by increasing the activities of anti-oxidative enzymes and decreasing the levels of malondialdehyde and total peroxide accumulation. Moreover, priming also enhanced the anti-oxidative activities of treated seeds. The activities of isocitrate lyase and malate synthase were also increased in primed seeds. The enhanced anti-oxidation and lipid-carbohydrate conversion activities might explain in part why primed purple coneflower seeds emerged better than non-primed seeds.  相似文献   

3.
Seed balls of ‘Early Wonder Tall Top’ table beet (Beta vulgaris L.) were incubated for 2 days at 21 °C in moist exfoliated grade 3 vermiculite (3 g seed balls [168 seed balls] and 1.1 g vermiculite) containing equal weights of Trichoderma harzianum Rifai strain KRL-AG2 G41 and T. virens strain G-41 (ThTv) at 0, 0.25, 0.50, 0.75 or 1.00 mg ThTv per seed ball. ThTv also was applied to the peat-lite growth medium 14 days before planting, at the same rates per seed ball as the seed ball treatments. Four days before planting, the peat-lite was inoculated with Pythium aphanidermatum (Edson) Fitzp. (Pa) at 0, 0.5 and 1.0× the rate that resulted in 96% damping-off when non-ThTv-treated dry seed balls were sown in peat-lite containing 1.0 Pa. Increasing ThTv level per seed ball decreased percentage damping-off, with 1.0 Pa giving greater percentage damping-off than 0.5 Pa. At 1 mg ThTv per seed ball, damping-off was 5% and 19% at 0.5 and 1.0 Pa, respectively. Including Agro-Lig UF (mostly humic acids) in the incubating seed-ball-ThTv mixture further decreased damping-off by an average 13 and 10 percentage points in 0.5 and 1.0 Pa, respectively. Increasing ThTv per seed ball in growth media decreased percentage damping-off, but not to the extent achieved with seed ball treatment, with 1 mg ThTv per seed ball giving 20% and 55% damping-off in 0.5 and 1.0 Pa, respectively. Decreasing incidence of damping-off with increasing ThTv application to seed balls or growth media was associated with increasing shoot fresh weight m−2 at 14 days after planting, a response attributable to increased percentage plant survival and not to a ThTv growth-promoting effect.  相似文献   

4.
The bitter gourd seed has a thick, hard seed coat. Mildew often occurs during germination and causes uneven and low rates of seed germination. However, the problems caused by mildew can be overcome by treating seeds with hot water, by soaking in water, or by using microorganisms. Seeds of the ‘Ching Pi’ bitter gourd were treated in a water bath at 60 °C for 10 min and then soaked in tap water at 25 °C for 24 h. The resulting germination percentage was 86.7%, and the resulting percentage of mildewed seeds was 10%. The biocontrol potential of three commercially available Bacillus subtilis solutions was examined. For seeds primed with Huodijun B. subtilis solution, the germination percentage was 73.3% and the mildewed percentage 6%. In dual cultures, the antibiotic content in the Huodijun B. subtilis solution was significantly greater than in Yunghsing and Huolibao, the other B. subtilis solutions examined. B. subtilis effectively inhibited the growth of Rhizoctonia solani mycelium and caused abnormal mycelial growth.  相似文献   

5.
To develop efficient seedling production methods for Laccosperma secundiflorum and Eremospatha macrocarpa, a study was conducted to examine regeneration using offsets combined with several physical and chemical treatments of seeds. Offsets categorized into small, medium and large diameters, were planted in three conditions: shaded and open nursery, and greenhouse. We tested sucker from E. macrocarpa, and sucker and rhizome from L. secundiflorum. For both species, high viability percentage (ranging from 55% to 100%) were observed for small and medium suckers planted in shaded nursery and greenhouse, against less than 49% for sucker planted in open nursery. The mean seedling emergence times were estimated to 84, 77 and 75 days after planting (DAP) for small, medium and large sucker of L. secundiflorum, respectively under open nursery condition, and 76, 75, 95 DAP for small, medium and large suckers of the same species, respectively in shaded condition. Greenhouse has a significant positive effect on E. macrocarpa seedlings emergence time. For this species, the mean seedling emergence times were estimated to 43 DAP for small sucker and 76, 93 DAP for medium and large suckers. No seedling was obtained from rhizome planted in all the growing conditions tested. Concerning seed dormancy breaking, germination percentages and rates were determined for 13 treatments. The best treatments were pre-soaking unscarified seeds for 4 days in 1.01 g l−1 and 0.10 g l−1 KNO3, with 79% and 68% of germination, respectively and in 3.46 × 10−3 g l−1 GA3 for 68% of germination. These methods are suggested to improve germination of L. secundiflorum seeds. Successful and recommended methods for E. macrocarpa are pre-soaking scarified seeds in 3.46 × 10−3 g l−1 and 3.46 × 10−4 g l−1 GA3, 96% and 94% of germination, respectively. Dormancy, probably a combination of mechanical and chemical dormancy, is present in the two species.  相似文献   

6.
A study was carried out to investigate the influence of desiccation and freezing followed by various presowing rehydration procedures on the desiccation sensitivity of the seed of Citrus suhuiensis cv. limau madu. The freshly harvested seeds of limau madu were desiccated under a broad range of relative humidity (RH) to various equilibrium water contents (g H2O g−1 dw). The desiccated and desiccated–frozen seeds were either directly sown under germination conditions or subjected to presowing rehydration procedures: seed preheating, prehumidification and osmoconditioning. The hydrated and desiccated seeds were sown in controlled germination conditions and the survival was evaluated 4–6 weeks after sowing. The results showed that desiccation progressively reduced the percentage of normal seedling of the seeds of limau madu and the viability is almost lost at water contents below 0.08 g H2O g−1 dw. The estimated desiccation sensitivity was substantially high (WC50 = 0.143 g H2O g−1 dw) when the desiccated seeds were rapidly rehydrated (uncontrolled rehydration). In contrast, seed prehumidification, preheating and osmoconditioning (controlled rehydration procedures) markedly enhanced normal seedling percentages decreasing the estimated values of WC50 (between 0.08 and 0.127 g H2O g−1 dw). While the rapidly rehydrated desiccated–frozen seeds were almost killed at water content of 0.15 g H2O g−1 dw, prehumidification and preheating have noticeably increased percentage of frozen seeds survival at the same water content. However, at water content of 0.21 g H2O g−1 dw preheating significantly (P < 0.05) increased percentage of normal seedling of the frozen seeds. Seed desiccation markedly reduced the percentages of germinated seeds with multiple seedlings. Seed controlled rehydration remarkably increased the survival of polyembryos. The beneficial effect of seed controlled rehydration on the survival of the desiccated seeds was pronounced at medium water contents (0.08–0.25 g H2O g−1 dw).  相似文献   

7.
Michelia yunnanensis Franch. is a Chinese endemic ornamental shrub with potential for greater utilization as a landscape and medicinal plant if propagation was less difficult. Seed development and breaking of seed dormancy were investigated to improve propagation of M. yunnanensis. No fresh seeds germinated when tested at the time of dispersal. Newly matured seeds of M. yunnanensis contained differentiated linear underdeveloped embryos that were physiologically dormant. The embryo/seed length ratio of M. yunnanensis was 0.15. Warm stratification did not break seed dormancy. Dormancy was broken by cold stratification at 4 °C but not by flowing water or nitrate. Embryos developed grew inside seeds during cold stratification at 4 °C. In newly harvested dormant seeds, embryos were 0.94 mm long and increased in length 139% before radicle emergence (germination). GA3 substituted for cold stratification to break dormancy in seeds of M. yunnanensis incubated at 25 °C or 20/25 °C. Mature M. yunnanensis seeds exhibited intermediate complex morphophysiological dormancy. Optimal germination of non-dormant seed in terms of both germination percentage and rate occurred at 20/25 °C.  相似文献   

8.
The dormancy mechanisms of Areca triandra Roxb. Ex Buch-Ham seeds were studied by treating the intact or mechanically scarified seeds with scarification, chemical soaking and stratification. The results indicate that the seeds have exogenous and endogenous dormancy. The exogenous dormancy is imposed by the pericarp and it is the major limiting factor for germination. It can be broken by mechanical scarification, but not by chemical scarification in 98% H2SO4 for 30 min. Chemical treatments (soaking for 24 h in 100–200 mg/L GA3, 0.2% KNO3 and 0.1–0.3% NaNO2, and for 12 h in 10% H2O2 or 20 min or 12 h in 15% H2O2) and stratifications, especially, cold stratifications for 30–120 days, broke the endogenous dormancy and significantly hastened germination of mechanically scarified seeds, although they did not increase the germination percentages.  相似文献   

9.
Pre-dehiscent capsules were collected from two Rhododendron griersonianum (Balf.f. & Forrest) trees and either immediately dried in a dry-room (15% relative humidity, 15 °C) or placed in a high humidity room (80% relative humidity, 15 °C) for 30, 60, or 90 d. Further capsules were also collected from the trees at 30 and 60 d, but seeds had been dispersed by 90 d. Seed ageing experiments (60% relative humidity, 45 °C) carried out on these seed-lots and on seeds from a further 10 Rhododendron (L.) species confirmed that short seed lifespans is a trait of the genus, with a mean P50 value of ca. 20 d for this storage environment.  相似文献   

10.
Data on the growth-promoting effects of Azospirillum on lettuce exposed to either normal or saline conditions, is scarce. Lactuca sativa L., cv Mantecosa seeds were colonized with A. brasilense Sp245 cells during imbibition. Germination percentages were determined after 7 d treatments with 0, 30, 50 or 80 mol m−3 NaCl. In another experiment, seeds germinated in Hoagland were irrigated for 30 d with 0, 30, 50 or 80 mol m−3 NaCl supplemented media. Vegetative growth proceeded in a growth chamber with a 13–11 h day–night cycle. Buffer-imbibed seeds were considered non-inoculated controls. Plant samples were taken at 0, 14, 20, and 30 d after the onset of NaCl treatments and dissected in aerial and root portions. The weights of both tissues were measured. Azospirillum-inoculated seeds had significantly higher germination percentages than controls in all treatments. Inoculated dried seeds stored up to 30 d maintained such characteristic in most of the treatments, particularly at 80 mol m−3 NaCl. Plants grown from inoculated seeds and irrigated with saline media displayed higher total fresh and dry weights and biomass partition to the aerial portion, than non-inoculated controls. Azospirillum-inoculated lettuce seeds had better germination and vegetative growth than non-inoculated controls after being exposed to NaCl.  相似文献   

11.
This study was initiated to investigate the differences in germination percentages and rates between Corylopsis coreana Uyeki and Corylopsis sinensis var. calvescens Rehder & E.H. Wilson following a warm stratification (WS) and cold stratification (CS), and to study the effect of different WS temperatures interacting with different durations of CS. Warm stratification at 10 °C, 15 °C, 20 °C, and 25 °C was given for 1 month (1 M 10 °C, 15 °C, 20 °C, and 25 °C WS) followed by 0 M, 1 M, 2 M, and 3 M of CS at 5 °C (0 M, 1 M, 2 M, 3 M CS) and seeds were germinated in an air conditioned greenhouse maintained at 18.5 °C/18 °C. On average, less than 1% of C. coreana seeds germinated when sown without any WS and CS or with 1 M 15 °C, 20 °C, and 25 °C WS without CS treatment. However, 26% C. coreana seeds germinated after 1 M 10 °C WS without any CS treatment. Germination was not affected by WS temperatures when followed by 2 M 5 °C CS. It is concluded that C. coreana exhibited low seed germination at 10 °C and that this temperature could be considered the upper limit of CS for C. coreana. Only 2 M CS was required for more than 90% seeds to germinate. However, C. sinensis var. calvescens required longer than 3 M CS for more than 29% seeds to germinate. This clearly shows that there is an interspecific variation in optimum dormancy-breaking requirements.  相似文献   

12.
The present investigation was undertaken to develop PRSV (Papaya ringspot virus) resistant hybrids through intergeneric hybridization. Intergeneric hybridization was done involving nine Carica papaya cultivars as female and Vasconcellea cauliflora as male. To break the intergeneric hybridization barrier, various nutrient combinations were used. Among the combinations used, sucrose 5%, sucrose 5% + boron 0.5% and sucrose 5% + CaCl2 0.5% improved the fruit set and seed set percentage. A total number of 1197 flowers were pollinated and 308 fruits were obtained. On extraction, 721 seeds were obtained from CO 7, Pusa Nanha and CP 50. Out of 721 F0 seeds (crossed seeds) sown, 419 seeds germinated and artificial screening for PRSV was carried out 27 days after sap inoculation. Out of 29 F1 hybrid plants from CO 7 x V. cauliflora cross, only six plants namely CO 7V1 to CO 7V6 were found free from PRSV symptoms. Similarly, out of 55 F1 hybrids from cross involving Pusa Nanha x V. cauliflora only 23 plants namely PNV1 to PNV23 were found free from the symptoms and 70 plants namely CPV1 to CPV70 out of 335 plants of CP50 x V. cauliflora cross were found free from PRSV symptoms. Among the crosses, Pusa Nanha x V. cauliflora had higher yield under PRSV infected conditions, however, total soluble solids and total sugars were found lesser than the CO 7 x V. cauliflora cross. The hybridity of the progenies were confirmed by using ISSR (Inter Simple Sequence Repeats) primers by the amplification of DNA from progenies and their parents. ISSR primers UBC 856, UBC807 and ISSR primer combinations UBC 856-817, UBC 810-817, UBC 861-817, UBC 856-810, UBC 861-810 and UBC 856-817 clearly amplified specific bands of the male parent, which were present in F1 progenies, but it was absent in female parents.  相似文献   

13.
The objective of this study was to produce interspecific hybrids between an Ogura-cytoplasmic male sterile (CMS) line of zicaitai (Brassica campestris var. purupurea, 2n = 20) and cultivars of ornamental kale (Brassica oleracea var. acephala, 2n = 18) to develop a CMS system for hybrid seed production. Pollination with pollen grains of ornamental kales irradiated at a power output of 9.0 mW with a He–Ne laser for 3 min could overcome the cross-incompatibility between the species concerned. Intact hybrids could be efficiently produced from ovules cultured on Murashige and Skoog media supplemented by 0.2 mg l−1 6-benzyladenine. Chromosome number of hybrids was confirmed to be 2n = 19. Hybrids resembled ornamental kales in leaf morphology and in vernalization response. Pollens of hybrids had a sterile appearance. Moreover the hybridity of the putative hybrids was confirmed by RAPD data on a DNA fragment of 820 bp.  相似文献   

14.
Linaria maroccana Hook. f. Ann., ‘Lace Violet’, Lupinus hartwegii ssp. cruikshankii Lindl. ‘Sunrise’ and Papaver nudicaule L. ‘Meadow Pastels’ seeds were directly sown into 105 cell plug trays and received either ambient light or supplemental high intensity discharge (HID) lighting. For each species, a 2 × 3 × 3 factorial was used with two light intensities during propagation, three transplant stages, and three night temperatures. Seedlings were transplanted at the appearance of 2–3, 5–6, or 8–9 true leaves. Transplanted Linaria and Papaver seedlings were placed at 5/11, 10/16, or 15/21 ± 1 °C night/day temperatures and Lupinus seedlings were placed at 15/24, 18/25, or 20/26 ± 2 °C night/day temperatures. For this study, the optimum production temperature for Linaria was 10/16 °C as the cut stems produced at 15/21 °C were unmarketable and production time was excessively long at 5/11 °C. At 10/16 °C, Linaria seedlings should be transplanted at the 2–3 leaf stage to maximize stem number, stem length and profitability. For Lupinus the optimum temperature was 15/24 °C due to long stems and high profitability per plant. Lupinus seedlings should be transplanted at the 2–3 leaf stage when grown at 15/24 °C to obtain the longest and thickest stems; however, $/m2 week was higher for plants transplanted at the 8–9 leaf stage due to less time in finishing production space. For Papaver, the 15/21 °C temperature was optimal as that temperature produced the longest stems in the shortest duration, resulting in the highest $/m2 week. At 15/21 °C Papaver plants should be transplanted at the 2–3 leaf stage. Supplemental HID lighting had no effect on any of the species.  相似文献   

15.
The effects of different types of chitosan on seed germination and protocorm development were determined for two orchid species, Dendrobium bigibbum var. compactum and Dendrobium formosum. Six chitosan types derived from polymer or oligomer chitosan each with 70, 80 or 90% levels of deacetylation (P70, P80, P90, O70, O80 and O90, respectively), were evaluated as direct medium supplements at 0, 10, 20, 40 or 80 mg/L in modified VW medium by following seed germination and protocorm growth for 12 weeks. Chitosan of all six tested types and four concentrations were found to significantly enhance the proportion of D. formosum seeds that germinated, when compared to these germinated without chitosan. In contrast, chitosan caused no enhanced germination rate was noted for D. bigibbum var. compactum with all tested chitosans and doses tested. However, almost all types of chitosan at 10 mg/L, except O90, were able to significantly improve the growth of D. bigibbum var. compactum protocorms, whilst 10 or 20 mg/L of P70 chitosan was the best formula to enhance the growth of D. formosum protocorms. It is concluded that chitosan responses in seed germination and protocorm development were somewhat species and developmental stage dependent. Therefore, the appropriate chitosan application for each plant species should be evaluated first before use.  相似文献   

16.
Passiflora mollissima, Passiflora tricuspis and Passiflora nov sp. are three passion fruit species occurring in Bolivia. Germination percentages and rates were determined for 11 different treatments. Per species, germination of 100 seeds was monitored every 3 days, during 90 days. Germination started after 9 days and 50% of final germination was reached within a month or less. Successful, recommended methods for P. mollissima are removing the basal point of seeds (27% germination) or removing the basal point in combination with pre-soaking seeds for 48 h in 50 ppm GA3 (18%). Pre-soaking seeds for 24 h in 400 ppm GA3 (42%) and removal of the basal point in combination with pre-soaking seeds for 48 h in 50 ppm GA3 (36%) are suggested methods to improve germination of P. nov sp. Removing the apical point of P. tricuspis seeds resulted in maximal germination (57%). No unique treatment gave satisfactory results for the three species tested. Exogenous dormancy, probably a combination of mechanical and chemical dormancy is present in the three species studied. Presence of physical dormancy was found in P. mollissima.  相似文献   

17.
Dessert quality of guava fruit is considerably reduced by high seed content. Number of seeds in guava is associated with different seed and fruit characters. Influence of different seed characters on number of seeds per fruit (NSPF) was studied by analysing character association and direct and indirect effects on NSPF and seed related traits for 68 genotypes of guava collected from diverse sources and conserved at National Active Germplasm Site, CISH, Lucknow. Seed related characters were studied at the colour break stage of the fruits using image analysis software for counting number of seeds. The data was subjected to path analysis to find out direct and indirect effects of different characters on number of seeds in the fruits. At genotypic and phenotypic levels, NSPF was significantly and positively associated with seed weight per fruit (SWPF), number of seeds 100 g−1 pulp (NSPHP) and fruit weight (FW). The genotypic (0.0029) and phenotypic (0.0563) residual values were fairly very low, which revealed that variables included in this study had significant contribution in determining NSPF. The NSPHP exhibited high positive direct effect on NSPF. The maximum direct response (P = 0.737, G = 1.004) of this component was attributed to the indirect positive effects of the SWPF and fruit:seed weight proportion (FSWP). SWPF also had a very high positive direct effect (P = 0.521, G = 0.694) on the NSPF. Correlation and path coefficient analysis revealed that SWPF, NSPHP and 100-seed weight (100SW) were deciding factors for realizing improvement for NSPF. The importance of small seeded genotypes in selection of less seeded varieties was emphasized.  相似文献   

18.
Cyclocarya paliurus is native to China and is the sole species in its genus. However, the seeds remain deeply dormant for 2 years in their natural environment. We tested different pretreatments of chemical scarification and exogenous application of gibberellic acid (GA3) for efficacy in breaking dormancy and speeding germination. In contrast to scarified seeds, non-scarified seed did not germinate, indicating that C. paliurus seeds have hard, impermeable seed coat dormancy. Exogenous application of GA3 significantly enhanced germination of scarified seeds. Compared with seeds stratified in sand with water, the germination of seeds stratified in sand moistened with 400 ppm GA3 for 60 days was significantly increased and germination rate was over 90% after 120 days. Analysis of variance indicated that both GA3 concentration and stratification medium had significant effects on seed germination and final germination percentage. Germination was higher for longer stratification periods, but no significant difference in germination was observed after 90 days. These results suggested that C. paliurus seeds exhibit both exogenous and endogenous dormancy. A combination of chemical scarification and exogenous application of GA3 alleviated seed dormancy in a relatively short period of time.  相似文献   

19.
Mal secco (caused by Phoma tracheiphila (Petri) Kantsch. and Gik.) is the most destructive fungal disease of lemon plantations worldwide and seedless lemons would be preferred by most consumers. Five dosage levels, 0, 3, 5, 7, and 9 kiloradian (krad), of cobalt (60Co) gamma irradiation were applied to budstick of ‘Kutdiken’ lemon (Citrus limon (L.) Burm. f.) clone KT-2A. Mutations were stabilized in three vegetative generations. Three hundred fifty-eight and 478 M1V3 (mutation one and vegetation three) plants were evaluated for seed number and mal secco tolerance in the field and the greenhouse, respectively. LD50 was around 5 krad gamma irradiation for ‘Kutdiken’ lemon. The seed number varied from 0 to 34 per fruit. The level of mal secco tolerance also varied significantly among the plants from 1.0 (no symptom) to 4.3 (high level of disease occurance). The stable seedless and mal secco tolerant plants were obtained from 5 and 7 krad irradiation: the three mutants from 5 krad irradiation gave more lemon-like fruits, while 7 krad irradiation caused altered tree morphology and early maturation of fruits. This study shows considerable potential for lemon cultivar improvement aiming to obtain seedless and mal secco tolerant lemons.  相似文献   

20.
A protocol is developed for Agrobacterium-mediated genetic transformation of Amaranthus tricolor via explant co-cultivation with Agrobacterium rhizogenes. Bacteria-plant specific factors which influenced transformation were optimized. Of the two Agrobacterium strains employed, LBA9402 was more infectious compared to A4. Bacterial suspensions grown overnight with 100 μM acetosyringone and experiencing O.D.660 = 0.6 followed by dilution to a density of 109 cells ml−1 were the most effective. Explants from garden-grown plants were more responsive than those from in vitro cultures; stem internodes being better than leaves. Immersion of the pre-pricked explants in bacterial suspension resulted in a markedly higher transformation frequency compared to the direct injection method. The infection of internode explants with the LBA9402 strain followed by co-cultivation on growth regulator-free MS medium (MS0) for 5 days resulted in emergence of hairy roots up to a maximum frequency of 97.22%. Roots were individually cultured in MS0, but fortified with bactericidal antibiotic (500 μg ml−1 cefotaxime). Rhizoclones showing prolific growth were renewed through successive subcultures in MS0. Opine gene expression was revealed by positive agropine and mannopine synthesis in all selected transformed rhizoclones. Shoot regeneration from root clones, capable of auxin-independent growth and opine proficiency, was stimulated in MS augmented with 2.0 mg l−1 zeatin. pRi TL–DNA rolB and pRi TR–DNA man2 ORF were detected in leaf tissues of regenerated plants from selected hairy root clones through PCR amplification. The implication of such findings is discussed on the possibility of conferring protection to crop amaranths against biotic stress challenges, particularly due to insects, viruses or fungal pathogens.  相似文献   

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