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1. Genetic and agronomic traits stability of marker-free transgenic wheat plants generated from Agrobacterium-mediated co-transformation in T2 and T3 generations
LIU Hui-yun, WANG Ke, WANG Jing, DU Li-pu, PEI Xin-wu, YE Xing-guo
Journal of Integrative Agriculture    2020, 19 (1): 23-32.   DOI: 10.1016/S2095-3119(19)62601-8
摘要99)      PDF    收藏
Genetically modified wheat has not been commercially utilized in agriculture largely due to regulatory hurdles associated with traditional transformation methods.  Development of marker-free transgenic wheat plants will help to facilitate biosafety evaluation and the eventual environmental release of transgenic wheat varieties.  In this study, the marker-free transgenic wheat plants previously obtained by Agrobacterium-mediated co-transformation of double T-DNAs vector were identified by fluorescence in situ hybridization (FISH) in the T1 generation, and their genetic stability and agronomic traits were analyzed in T2 and T3 generations.  FISH analysis indicated that the transgene often integrated into a position at the distal region of wheat chromosomes.  Furthermore, we show that the GUS transgene was stably inherited in the marker-free transgenic plants in T1 to T3 generations.  No significant differences in agronomic traits or grain characteristics were observed in T3 generation, with the exception of a small variation in spike length and grains per spike in a few lines.  The selection marker of bar gene was not found in the transgenic plants through T1 to T3 generations.  The results from this investigation lay a solid foundation for the potential application of the marker-free transgenic wheat plants achieved through the co-transformation of double T-DNAs vector by Agrobacterium in agriculture after biosafty evaluation.
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2. Genetic behavior of Triticum aestivum–Dasypyrum villosum translocation chromosomes T6V#4S·6DL and T6V#2S·6AL carrying powdery mildew resistance
LIU Chang, YE Xing-guo, WANG Mei-jiao, LI Shi-jin, LIN Zhi-shan
Journal of Integrative Agriculture    2017, 16 (10): 2136-2144.   DOI: 10.1016/S2095-3119(16)61568-X
摘要628)      PDF    收藏
T6V#2S·6AL and T6V#4S·6DL translocation chromosomes developed from the cross of wheat and different Dasypyrum villosum accessions have good powdery mildew (PM) resistance, but their pairing and pyramiding behavior remains unclear.  Results in this study indicated that the pairing frequency rate of the two differently originated 6VS chromosomes in their F1 hybrid was 18.9% according to genomic in situ hybridization (GISH); the PM resistance plants in the F2 generation from the cross between T6V#4S·6DL translocation line Pm97033 and its PM susceptible wheat variety Wan7107 was fewer than expected.  However, the ratio of the resistant vs. the susceptible plants of 15:1 in the F2 generation derived from the cross between the two translocation lines of T6V#2S·6AL and T6V#4S·6DL fitted well.  Plants segregation ratio (homozygous:heterozygous:lacking) revealed by molecular marker for T6V#4S·6DL or T6V#2S·6AL in their F2 populations fitted the expected values of 1:2:1 well, inferring that the pairing of the two alien chromosome arms facilitates the transmission of T6V#4S·6DL from the F1 to the F2 generation.  A quadrivalent was also observed in 21% of pollen mother cells (PMCs) of homozygote plants containing the two pairs of translocated chromosomes.  The chromosome pairing between 6V#2S and 6V#4S indicates that it will be possible to obtain recombinants and clarify if the PM resistance determinant on one alien chromosome arm is different from that on the other.  
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3. A genetic evidence of chromosomal fragment from bridge parent existing in substitution lines between two common wheat varieties
ZHAO Pei, WANG Ke, LIN Zhi-shan, LIU Hui-yun, LI Xin, DU Li-pu, YAN Yue-ming, YE Xing-guo
Journal of Integrative Agriculture    2016, 15 (1): 10-17.   DOI: 10.1016/S2095-3119(15)61038-3
摘要1845)      PDF    收藏
Locating of important agronomic genes onto chromosome is helpful for efficient development of new wheat varieties. Wheat chromosome substitution lines between two varieties have been widely used for locating genes because of their distinctive advantages in genetic analysis, compared with the aneuploid genetic materials. Apart from the substituted chromosome, the other chromosomes between the substitution lines and their recipient parent should be identical, which eases the gene locating practice. In this study, a set of chromosome substitution lines with cv. Wichita (WI) as the recipient parent and cv. Cheyenne (CNN) as the donor parent were studied for the composition of high molecular weight glutenin subunits (HMW-GS) as well as a range of agronomic important traits. Results revealed that the substitution lines of WI(CNN5D), WI(CNN6A) and WI(CNN7B) had higher plant heights than the two parents of WI and CNN, and WI(CNN3D) had later maturity than the parents. By sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) analysis, a substitution line WI(CNN5B) was found to contain different HMW-GS patterns from its two parents, in which 1By9 was replaced by 1By8 on chromosome 1BL. Simple sequence repeat (SSR) analysis confirmed that the variation on 1BL in WI(CNN5B) was originated from Chinese Spring (CS). It is concluded that chromosomal fragments from bridge material and donor parent were quite often retained in intracultivaral chromosome substitution lines except the substituting chromosomes.
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4. Current status of genetic transformation technology developed in cucumber (Cucumis sativus L.)
WANG Shun-li, Seong Sub Ku, YE Xing-guo, HE Cong-fen, Suk Yoon Kwon, Pil Son Choi
Journal of Integrative Agriculture    2015, 14 (3): 469-482.   DOI: 10.1016/S2095-3119(14)60899-6
摘要2044)      PDF    收藏
Genetic transformation is an important technique for functional genomics study and genetic improvement of plants. Until now, Agrobacterium-mediated transformation methods using cotyledon as explants has been the major approach for cucumber, and its frequency has been up to 23%. For example, significantly enhancement of the transformation efficiency of this plant species was achieved from the cotyledon explants of the cultivar Poinsett 76 infected by Agrobacterium strains EHA105 with efficient positive selection system in lots of experiments. This review is to summarize some key factors influencing cucumber regeneration and genetic transformation, including target genes, selection systems and the ways of transgene introduction, and then to put forward some strategies for the increasing of cucumber transformation efficiency. In the future, it is high possible for cucumber to be potential bioreactor to produce vaccine and biomaterials for human beings.
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5. Development and application of plant transformation techniques
YE Xing-guo
Journal of Integrative Agriculture    2015, 14 (3): 411-413.   DOI: 10.1016/S2095-3119(14)60945-X
摘要2279)      PDF    收藏
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6. Effects of inter-culture, arabinogalactan proteins, and hydrogen peroxide on the plant regeneration of wheat immature embryos
ZHANG Wei, WANG Xin-min, FAN Rong, YIN Gui-xiang, WANG Ke, DU Li-pu, XIAO Le-le, YE Xing-guo
Journal of Integrative Agriculture    2015, 14 (1): 11-19.   DOI: 10.1016/S2095-3119(14)60764-4
摘要1906)      PDF    收藏
The regeneration rate of wheat immature embryo varies among genotypes, howbeit many elite agriculture wheat varieties have low regeneration rates. Optimization of tissue culture conditions and attempts of adding signal molecules are effective ways to increase plant regeneration rate. Inter-culture is one of ways that have not been investigated in plant tissue culture. Moreover, the use of arabinogalactan proteins (AGPs) and hydrogen peroxide (H2O2) have been reported to increase regeneration rate in a few plant species other than wheat. The current research pioneeringly uses inter-culture of immature embryos of different wheat genotypes, and also investigates impacts of AGP and H2O2 on the induction of embryogenic calli and plant regeneration. As a result, high-frequency regeneration wheat cultivars Kenong 199 (KN199) and Xinchun 9 (XC9), together with low-frequency regeneration wheat line Chinese Spring (CS), presented striking increase in the induction of embryogenic calli and plant regeneration rate of CS through inter-culture strategy, up to 52.19 and 67.98%, respectively. Adding 50 to 200 mg L–1 AGP or 0.005 to 0.01 ‰ H2O2 to the callus induction medium, enhanced growth of embryogenic calli and plant regeneration rate in quite a few wheat genotypes. At 50 mg L–1 AGP application level in callus induction medium plant regeneration rates of 8.49, 409.06 and 283.16% were achieved for Jimai 22 (JM22), Jingdong 18 (JD18) and Yangmai 18 (YM18), respectively; whereas at 100 mg L–1 AGP level, CS (105.44%), Chuannong 16 (CN16) (80.60%) and Ningchun 4 (NC4) (62.87%) acted the best. Moreover CS (79.05%), JM22 (7.55%), CN16 (101.87%), YM18 (365.56%), Yangmai 20 (YM20) (10.48%), and CB301 (187.40%) were more responsive to 0.005 ‰ of H2O2, and NC4 (35.37%) obtained the highest shoot regeneration rates at 0.01 ‰ of H2O2. Overall, these two methods, inter-culture and AGP (or H2O2) application, can be further applied to wheat transgenic research.
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