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1.
To determine the most effective dose of arabinogalactan-protein (AGP) in regeneration medium, mature embryos of genotypes in three different ploidy levels (Triticum aestivum L. cv. Ikizce-96, Triticum durum Desf. cv. Mirzabey and Hordeum vulgare L. cv. Tokak) were used to establish an efficient plant regeneration system for cereals. Percentage of callus production, capacity of regeneration were calculated, and also culture effect, root, stem, and total plant length of regenerant plants were observed in six different regeneration media (MS control, MS+2, 5, 7, 10, 12 mg L-1 AGP) in these three different genotypes. According to the results, the highest amount of callus production was found in Ikizce-96 as 93.75% using 5 mg L-1 dicamba and 1 mg L-1 kinetin in induction medium. However, the most improved callus was observed in diploid barley Tokak as 179.95 mg in weight and 6.18 mm in diameter, respectively. The highest regeneration capacity was observed in the dose of 5 mg L-1 AGP in MS of all the genotypes and hexaploid wheat Ikizce-96 gave the best results with the highest regeneration capacity and culture effects (94.86 and 92.5%) in the same dose of AGP. These results indicated that effective dose of AGP in regeneration medium increase plant regeneration in calli derived from cereal mature embryos.  相似文献   

2.
The F1 and F4 plants of ‘synthetic hexaploid wheat/common wheat’ crosses and part of their parents were inoculated with Fusarium graminearum to evaluate FHB resistance.The results showed tht the scab resistance in the F1 varied with the synthetic wheat accessions used as crossing parents.In the F4,some resistant head lines were generated from the crosses,although their parents had different scab resistance levels.It indicated that synthetic hexaploid wheat are useful in wheat breeding for scab resistance.  相似文献   

3.
The haploid-inducing line Stock 6 was used to produce haploid maize and expected to obtain maize haploid plants successfully. The detailed meiotic studies on selected haploid maize （n = x = 10） were conducted. Cytogenetic analysis revealed a high frequency of meiotic abnormality occurred in both meiosis Ⅰ and meiosis Ⅱ. During the prophase Ⅰ, univalents were common configurations, and there were bivalents or trivalents in some pollen mother cells, however, a few cells containing five bivalents were also observed. After prophase Ⅰ, chromosomes did not congregate in a single metaphase plate but they were scattered in the cytoplasm. At anaphase Ⅰ, the chromosome distribution was highly irregular with almost all possible combinations. In some cells, chromosomes were grouped into the three or four masses and several spindles appeared. At the tetrad stage of meiosis Ⅱ, eytokinesis splitting abnormality occurred, and a variety of diad, triad, tetrad, pentad, hexad, as well as decury microspores were easily observed. As a consequence of abnormalities of the two meiotic stages, various microspores and the pollen were almost completely sterile. grains with different size were formed, and its pollen grains  相似文献   

4.
Wheat is the number one crop both in acreage and total yield in the world. Therefore, it is very important to improve wheat by gene engineering techniques even though it belongs to the plants insensitive to gene transformation, especially to Agrobacterium-mediated transformation. Wheat immature embryos of 1.0-1.5mm in size, C58c1 of Agrobacterium strain harboring pPTN249, pPTN270, pPTN254, and pSIS-GFP respectively (all the vectors contain the aphA selectable gene driven by enhanced 35S promoter and a target gene controlled by ubi promoter or E35S promoter), AB medium for Agrobacterium activate culture, WCC medium for co-culture, were used in this study. The embryos with 4 days of pre-culture were transferred onto selection medium with 10mg/L geneticin, 50mg/L ticarcillin, 50mg/L vancomycin, and 50mg/L cefotaxine after 30 minutes of infection and 2 days of co-cultivation with Agrobacterium. Followed callus production,shoot regeneration on selection medium, 114 resistant plantlets were obtained from 10 transformation experiments of four genotypes. By nptⅡ ELISA (nptⅡ enzyme assay), PCR, Southern blot and leaf bleach,29 positive plants were identified from two genotypes of Bobwhite and Yanglmai 10, with an average transformation efficiency of 0.82 %. The result tested by Southern blot also showed that the transgenic plants with single- copy integration of target gene took 65.52% among total positive plants. The ELISA value of transgenic plant was also related to the copies of alien DNA integrating into wheat chromosomes, the transgenic plants with single copy integration giving higher ELISA value than the ones with 2 or 3 copies integration.  相似文献   

5.
[Objective] The experimental was aimed to screen the optimum regeneration shoot induction media and rooting media for tender leaves of Rhododendron chrysanthum Pall.[Method] The tender leaves of Rhododendron chrysanthum Pall were taken as explants to select the optimum bud induction media and rooting media through uniform design and the screening results were verified.[Result] The optimum media for regeneration shoot of Rhododendron chrysanthum Pall contained 1/4 MS,3.70 mg/L ZT, 0.02 mg/L IAA and 1.00 mg/L KT and its induction rate was 95.5% and the rooting media contained modified MS, 0.10 mg/L IAA and 0.07 mg/L NAA and its rooting rate was 98%. [Conclusion] Through this experiment, regeneration systems for regeneration shoot and regenerated plant from tender leaves of Rhododendron chrysanthum Pall were created successfully.  相似文献   

6.
Selection of primary trisomics of the cabbage （Brassica oleracea var.capitata L） forms an important basis for gene chromosome mapping and for other genetic studies. The cabbage self-fertilization line - 9601 was used as material, using the root-tip cell chromosome number and pollen mother cell chromosome number identification and karyotype analysis to select the primary trisomics from the progenies of 3x x 2x in the cabbage. Many aneuploid plants with one or two extra chromosomes were obtained and a set of primary trisomics （Tri-1, Tri-2, Tri-3, Tri-4, Tri-5, Tri-6, Tri-7, Tri-8, and Tri-9, in which the Tri-1 and Tri-4 were from 2n＋2 plants and others from 2n＋ 1 plants） was acquired from these plants. Each trisomic exhibited some unique features, such as plant height, plant type, leaf type, size of flower bud, and inflorescence. The triploid crossing by the diploid is a convenient and effective way to select trisomics in the cabbage.  相似文献   

7.
An efficient plant regeneration system was developed from the immature embryos of Triticum aestivum L. Thinopyrum intermedium alien disomic addition lines, which resistant to powdery mildew. The protocol was based on a series of experiments involving the callus induction and differentiation. The experiment studied the effects ofmature embryos. We found that the embryembryo size on callus induction and differentiation of the immature embryos. We found that the embryo size is critical for the establishment of embryogenic callus. Immature embryos (0.8 ~ 1.5 ram) showed high ability to produce embryogenie callus capable of regenerating green plants. The medium Murashige and Skoog‘ s (MS) added with 2mg/L 2, 4-diehlo-rophenoxyacetic acid (2, 4-D) gave the best embryogenic callus induction, maintenance and regeneration. The embryogenic callus maintained high regeneration during six subcultures in the callus induction medium. Suitable time ofpartialdesiccation could effectively improve the regeneration capacity of the callus cultured for 3-4 month. Bud green spot and root green spot were observed during the differentiation of callus and the difference between them was described. Regenerated shoots were rooted on half-strength MS medium containing 0.2 mg/L Naphthalene acetic acid (NAA). Plants were successfully transferred to soil and grew well. This efficient plant regeneration system provides a foundation for the study of somaclonal variation of Triticum aestivum L.- Thinopyrum intermedium alien disomic addition lines.  相似文献   

8.
泥胡菜的组织培养及高效无性系建立(英文)   总被引：1，自引：0，他引：1  
[Objective] The research aimed to study the rapid propagation technology and establish effective clone of Hemistepta lyrata Bunge. [Method] With tender stem of Hemistepta lyrata Bunge as material, the conditions needed in calluses induction and differentiation, adventitious bud differentiation and radication, test tube seedling cutting and transplantation were studied. [Result] The results showed that the optimum medium for granulated calluses induction from tender stem was MS+BA 0.3 mg/L+2,4-D 1-1.5 mg/L, for granulated calluses and adventitious bud differentiation was MS+AgNO3 1.5 mg/L +BA 0.4 mg/L +NAA 0.1 mg/L. 1/2 MS+IAA 0.6 mg/L was suitable for test tube seedling rooting and regeneration, and cinder was used as transplantation and cutting substrate. [Conclusion] This study will provide the scientific reference for choosing the feasible medium in tissue culture of Hemistepta lyrata Bunge.  相似文献   

9.
To establish a highly efficient plant regeneration system for wheat genetic transformation, the effects of three different concentrations of dicamba and two different sugar types on callus induction and plant regeneration from mature embryo cultures were evaluated. Callus induction and plant regeneration were obtained from mature embryos of two commercial cultivars Zhoumai 18 and Yumai 34 （Triticum aestivum L.） cultured on L3 basal medium. The results showed that the efficiency of mature embryo culture was significantly influenced by the genotypes, sugar types and dicamba concentrations. 4 mg L^-1 dicamba proved the best effective for inducing embryogenic callus and also gave the highest proportion of plants regenerated across the two cultivars. Substitution of maltose by sucrose significantly improved the plant regeneration efficiency in both cultivars. There was a significant interaction between genotype-by-sugar types, and sugar types-bydicamba concentrations. Overall, Zhoumai 18 gave the highest frequency of plant regeneration （82.65%） when dicamba concentration was 4.0 mg L^-1 and with sucrose in initial callus induction. These results will facilitate genetic transformation work with elite wheat.  相似文献   

10.
An in vitro shoot regeneration procedure was developed in pepper (Capsicum annuum L.) cytoplasmic male sterility (CMS) lines 9704A and 8214A using cotyledon as explant. The callus and bud cluster derived from cotyledon tissue explants were proliferated on Murashige and Skoog (MS) medium supplemented with different combinations of 6-benzladenine (6-BA), indole-3-acetic acid (IAA), gibberellic acid (GA3) and silver nitrate (AgNO3). From the formula of MS appended with 5.0 mg/L 6-BA, 1.0 mg/L IAA and 5.0 mg/L AgNO3, for the explants callus and bud cluster, the maximum differentiation rates (respectively 100.0% and 58.3%) and average number of adventitious bud from each explant (respectively 18.8 and 13.2) were obtained. The optimum medium combination for the elongation of adventitious bud was determined to be: MS+ 3.0 mg/L 6-BA + 1.0 mg/L IAA+ 5.0 mg/L AgNO3+ 2.0 mg/L GA3, from which the elongation rates of buds from callus and bud cluster were both 100%, and the average number of per explant adventitious bud number reached 6.3 and 5.8, respectively. And all the elongated shoots were successfully rooted on half-strength MS medium supplemented with 0.3-0.5 mg/L IAA.  相似文献   

11.
Vitis vinifera 87-1 plants infected by grapevine fleck virus(GFkV) and grapevine rupestris stem pitting-associated virus(GRSPaV) were used as the plant materials for virus elimination treatment. This study evaluated the effects of ribavirin at different concentrations(15 and 25 μg mL~(–1); R15 and R25, respectively), thermotherapy(37°C; T), and the combination of ribavirin and thermotherapy(R15+T and R25+T) on eliminating viruses from grapevine plants in vitro. Both R15 and R25 had phytotoxic effects and weakened plant growth. Thermotherapy positively affected the growth of grapevine plants. Plant height was significantly greater in T, R15+T, and R25+T than in CK, R15 and R25. The proportion of dead plants after T, R15+T, and R25+T was 51.4, 11.4, and 8.6%, respectively. The survival rates of regenerated plants after all treatments were 68.0%. Ribavirin concentration and treatment time were related to the regeneration of shoot tips and elimination efficiencies of the two viruses. The survival rates of plants after R15+T for 30, 40, and 50 days were 97.3, 90.7, and 74.4%, respectively. The elimination rates of GRSPaV from plants in the three time quantum were 55.6, 84.6, and 93.8%, respectively. The elimination rate of GFkV was 23.9% higher in R25(35/44) than in R15(25/45), and that of GRSPaV was 7.0% higher in R25 than in R15. The combination of thermotherapy and chemotherapy was found to have a positive effect on the eradication of GFkV and GRSPaV, and R25+T for 50 days was able to completely eliminate the two viruses from in vitro grapevines.  相似文献   

12.
Study on the Factors Influencing the Efficiency of Wheat Transformation   总被引：1，自引：0，他引：1  
Wheat transformation efficiency is closely related to several factors such as receptor genotype, constructed plasmid and selection procedure after bombardment or co-cultivation. In our study, several kinds of antibiotics, which were normally used in plant transformation to the selection genes of nptⅡ, bar and hpt,were tested for the optimal concentrations for wheat transformation. The results showed that 25 - 50mg/L of geneticin (G418) was suitable for the selection of nptⅡ, kanamycin or neomycin was not suitable for use. 3 5mg/L of phosphinothricin (PPT) or biolaphos could be used for the selection of bar, 100 - 150mg/L of hygromycin for the selection of hpt. Yangmai 158 and Yangmai 10 with high tissue culture response and good agronomic characteristics were screened from 25 potential Chinese wheat cultivars. The concentration changing of selectable agent in selection medium was helpful to obtain enough regeneration plantlets with strong root system.  相似文献   

13.
Erianthus arundinaceus is an important, closely related genus of Saccharum officinarum L. It is therefore important to understand how the chromosomes are transmitted when it hybridizes with sugarcane. The hybrids and backcross progenies of S. officinarum and E. arundinaceus and their parents were used for Karyotype analysis and to study the law of chromosome transmission. The results showed that the somatic chromosome number of both of the E. arundinaceus Hainan92-105 and Hainan92-77 were 2n = 60 = 60sm, belonging to type 1 A, and the BC1 YC01-21 was 2n = 104 = 100m ＋ 4sm, belonging to type 2C. The other six tested clones belonged to type 2B. The both F1s YC96-66 and YC96-40 that originated from Badila （2n = 80 = 70m ＋ 10sm） with E. Arundinaceus were 2n = 70 = 68m ＋ 2sm, which suggests an n ＋ n transmission. The cross between YC96-66 （female parent） and CP84-1198 （male parent, 2n = 120 = 114m ＋ 6sm） also followed the same genetic law and the somatic chromosome number of their progeny, YC01-3 （2n = 105 = 95m ＋ 10sm）. The cross derived from YC96- 40 （female） and CP84-1198 （male）, YC01-21 had 2n = 104 = 100m ＋ 4sm chromosomes, following the same genetic law of n ＋ n. However, YC01-36 had 2n = 132 = 130m ＋ 2sm chromosomes, which suggests a 2n ＋ n chromosome transmission. It can be inferred that the inheritance of chromosomes was very complex in the BC1. The difference in chromosome number between clones was as high as 28. This could be explained by the 2n ＋ n transmission of chromosomes. In addition, as there was not be a regular number of haploids, this phenomenon is termed as disequilibrium hybridization.  相似文献   

14.
Marker-free GFP transgenic tobacco plants were constructed based on Cre/lox site-specific recombination system. A GFP gene was introduced into the tobacco genome using the Bar gene as a linked selectable marker flanked by recombination sites in a directed orientation. The Bar gene expression box was subsequently excised from the plant genome by a strategy of Cre gene retransformation. After removal of the Cre-NPT Ⅱ locus by genetic segregation through self-cross, plants that incorporated only the GFP transgene were obtained. Transgenic tobacco plants mediated by Agrobacterium tumefaciens were obtained, which resisted herbicide Basta and GFP expressed well, then the Cre gene was subsequently introduced into 5 plants of them, respectively, by retransformation. The leaf disks from Cre transgenic plants were used to test the resistance to Basta on the medium with 8 mg L-1 of PPT. The results showed that few discs were able to regenerate normally, and the excision at 76-100% efficiency depended on individual retransformation events. Evidence for a precise recombination event was confirmed by cloning the nucleotides sequence surrounding the lox sites of the Basta sensitive plants. The result indicated that the excision event in the recombination sites was precise and conservative, without loss or alteration of any submarginal nucleotides of the recombination sites. Bar gene excised plants were selfpollinated to allow segregation of the GFP gene from the Cre-NPT Ⅱ locus. The progenies from self-pollinated plants were scored for Kan senstivity, then the segregation of GFP gene from Cre-NPT Ⅱ locus in the Kan senstive plants were confirmed by PCR analysis subsequently. Hence, constructing marker-free transgenic tobacco plants by Cre/lox sitespecific recombination system was reliable, and the strategy presented here should be applicable to other plants for the construction of marker-free transgenic plants as well.  相似文献   

15.
The regeneration ability of four alfalfa （Medicago sativa L.） cultivars, Xinjiang Daye, Longdong, Gannong 1 and Gannong 3, was studied, and the effects of various cultivars, explant sources and medium recipes on regeneration were compared. The better callus forming frequency obtained from hypocotyls of Xinjiang Daye is 88.5% and regeneration frequency is 9.8% in our initial experiments. To further optimize regeneration system for genetic transformation, we therefore changed concentrations of plant growth regulators and supplemented with glutamine into callus-induction and shoot-regeneration media. Callus forming frequency and shoot differentiation frequency were increased to 100%. The time taken to generate transgenic plants （16 weeks） was shorter than that for previouse procedure （25 weeks） and regeneration frequency was promoted to 15.1%. The results show that addition of glutamine is particularly important for shortening period of regeneration and promoting regeneration frequency. For study of genetic transformation of alfalfa, Agrobacterium tumefaciens-mediated transformation of Xinjiang Daye was developed based on this optimized regeneration system. The plant expression vector carrying two glutamine synthetases （GS 1 and GS2） and △1-pyrroline-5-carboxylate synthetase （P5CS） gene was used for alfalfa in vitro transformation. Six transgenic alfalfa plantlets with resistance to PPT were obtained. The introduction of foreign genes into plants was assessed in the transformants by PCR analysis and Southern hybridizations.  相似文献   

16.
An interspecific hybrid F1 of Cucumis hystrix Chakr. x Cucumis sativus L. （NC4406） was used to establish the developmental sequence and to characterize the male and female gametophytes at cytological level for further understanding of the phylogenic relationship and the mechanism of fertility or sterility in the interspecific hybrid F1 The development of male and female gametophytes was studied through meiotic analysis and paraffin section observation technique, respectively. Meanwhile, the fertility level was assessed through hybrid F1 backcrossing to cultivated cucumber 4406. Variable chromosome configurations were observed in the pollen mother cells （PMCs） of hybrid F1 at metaphase Ⅰ , e.g., univalents, bivalents, trivalents, quadravalents, etc. At anaphase Ⅰ and Ⅱ, chromosome lagging and bridges were frequently observed as well, which led to the formation of polyads and only a partial number of microspores could develop into fertile pollen grains （about 23.3%）. Observations of the paraffin sections showed numerous degenerated and abnormal embryo sacs during the development of female gametophytes, and only 40% of the female gametophytes could develop into normal eight-nuclear megaspore. On an average, 22.8 and 6.3 seeds per fruit could be obtained from the reciprocal backcross. The interspecific hybrid F1 of C. hystrix x NC4406 was partially fertile; however, the meiotic behaviors of hybrid F1 showed a high level of intergenomic recombination between C. hystrix and C. sativus chromosomes, which indicated that it plays an important role for introgression of useful traits from C. hystrix into C. sativus.  相似文献   

17.
[Objective]As a mosquito-repelling ornamental plant,Pelargonium×Citrosum Vanleenii(P.× Citrosum Vanleenii)is hard to be acquired because of its hybrid background,the paper was to a new regeneration system of(P.× Citrosum Vanleenii).[Method] By studying the influence of plant growth regulators(PGRs)on explant type(leaves and petioles),the optimal combinations of PGRs to maximize SELSs(somatic embryo-like structure)and buds were established.[Result]0.2 mg/L NAA+1.0 mg/L BA was best for LS(leaves segments)and 0.2 mg/L NAA + 1.5 mg/L BAs was best for PS(petioles segments).Cultured plantlets were successfully acclimatized in soil where they grew normally without any morphological variation.Although both LS and PS were usable,the leaf was a better explant for induction of embryogenic calli,somatic embryo-like structures and buds.[Conclusion]This work offered a rapid and efficient system for plant regeneration of P.×Citrosum Vanleenii.  相似文献   

18.
An efficient in vitro regeneration system by direct organogenesis from mature nodal and internodal stem segments of Newhall navel orange (Citrus sinensis L. Osbeck) was developed. Illuminating conditions together with plant growth regulators affected the adventitious bud regeneration frequency and efficiency. The initial 15 d darkness inoculation is beneficial for the adventitious bud regeneration. The highest regeneration frequency (85.2%) and bud formation efficiency (3.7 per responsive internodal stem segment) were obtained in the media supplemented with 1.0 mg L^-1 BAP and 0.5 mg L^-1 NAA. ABA at 0.2 mg L^-1 positively affected the bud formation efficiency, which amounted to 8.5 buds per internodal segment in the presence of BAP at 1.0 mg L^-1 The adventitious shoots successfully rooted and were transferred to the soil.  相似文献   

19.
番茄内生细菌的分离及其促生活性研究(英文)   总被引：5，自引：1，他引：4  
[Objective] The study aimed to investigate the growth-promoting activities of endophytic bacteria from tomato plants.[Method]The endophytic bacteria isolated from different tissues of tomato plants were analyzed for the effects of their growth-promoting activities on the germination and growth of tomato plants.The bacteria with growth-promoting activity were preliminarily identified.[Result]Totally 59 endophytic bacterial strains were isolated from roots and stems of tomatoes,of which 4 showed significantly growth-promoting activity to germination and growth of tomato.The results suggest that these strains are endowed with the potential capability of growth-promoting.[Conclusion]The endophytic bacteria with growth-promoting activity were found among the isolates from tomato plants.This provided a good foundation for utilization of these bacteria with growth-promoting activity.  相似文献   

20.
番茄内生细菌的分离及其促生活性研究(英文)   总被引：4，自引：0，他引：4  
[Objective] The study aimed to investigate the growth-promoting activities of endophytic bacteria from tomato plants.[Method]The endophytic bacteria isolated from different tissues of tomato plants were analyzed for the effects of their growth-promoting activities on the germination and growth of tomato plants.The bacteria with growth-promoting activity were preliminarily identified.[Result]Totally 59 endophytic bacterial strains were isolated from roots and stems of tomatoes,of which 4 showed significantly growth-promoting activity to germination and growth of tomato.The results suggest that these strains are endowed with the potential capability of growth-promoting.[Conclusion]The endophytic bacteria with growth-promoting activity were found among the isolates from tomato plants.This provided a good foundation for utilization of these bacteria with growth-promoting activity.  相似文献   

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