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1.
旱作条件下转甜菜碱醛脱氢酶基因水稻籽粒灌浆特性的研究 总被引:1,自引:1,他引:1
在旱作条件下,对转甜菜碱醛脱氢酶(BADH)基因水稻的3个品系(52-7、51-15、51-22)、受体亲本中花8号(ck1)和当地推广的以色列陆稻白珍珠(ck2)的籽粒灌浆特性进行了比较分析。结果表明:5个品种(系)的强、弱势粒灌浆过程均符合Richards模型,强、弱势粒灌浆为异步灌浆型。三个转BADH基因品系与其受体亲本相比,强、弱势粒的灌浆速率提高,活跃灌浆期延长,灌浆高峰期的籽粒重增加;籽粒灌浆中、后期,强势粒的灌浆天数缩短,强、弱势粒的平均灌浆速率提高,且不同粒位籽粒灌浆速率差异减小,最终使不同粒位籽粒大小的差异缩小和粒重增加。最大灌浆速率、平均灌浆速率和灌浆高峰期的粒重与千粒重存在显著正相关。 相似文献
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Improvements of TKC Technology Accelerate Isolation of Transgene-Free CRISPR/Cas9-Edited Rice Plants
He Yubing Zhu Min Wang Lihao Wu Junhua Wang Qiaoyan Wang Rongchen Zhao Yunde 《水稻科学》2019,26(2):109-117
Elimination of the CRISPR/Cas9 constructs in edited plants is a prerequisite for assessing genetic stability, conducting phenotypic characterization, and applying for commercialization of the plants. However, removal of the CRISPR/Cas9 transgenes by genetic segregation and by backcross is laborious and time consuming. We previously reported the development of the transgene killer CRISPR (TKC) technology that uses a pair of suicide genes to trigger self-elimination of the transgenes without compromising gene editing efficiency. The TKC technology enables isolation of transgene-free CRISPR-edited plants within a single generation, greatly accelerating crop improvements. Here, we presented two new TKC vectors that show great efficiency in both editing the target gene and in undergoing self-elimination of the transgenes. The new vectors replaced the CaMV35S promoter used in our previous TKC vector with two rice promoters to drive one of the suicide genes, providing advantages over our previous TKC vector under certain conditions. The vectors reported here offered more options and flexibility to conduct gene editing experiments in rice. 相似文献
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单精注射法生产转基因小鼠的研究 总被引:1,自引:0,他引:1
精子胞质内显微受精技术(ICSI)作为辅助受精的一种手段,将体外受精研究和胚胎的显微操作技术结合起来,比以往对精子质量的要求大大降低,使其无论在畜牧业生产实践还是在哺乳动物的生殖生理基础研究中都有着十分重要的意义。本研究用小鼠精子及小鼠精子与GFP基因孵育后对小鼠卵母细胞进行ICSI,获得子代鼠。提取鼠尾基因组DNA,应用PCR、Southern blot进行整合检测。在发育至成年的11只小鼠中经PCR和Southernblot检测到3只阳性(27.3%)。结果表明,利用ICSI技术可以高效地生产转基因小鼠。 相似文献
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冷休克结构域蛋白(CSPs)是含有不同数量冷休克结构域(cold shock domain,CSD)的低温响应蛋白,在低温胁迫中发挥重要作用。以沙冬青(Ammopiptanthus mongolicus)为材料,克隆得到了冷休克结构域蛋白基因AmCSDP。生物信息学分析显示AmCSDP(Gen Bank登录号:KX756575)全长540bp,编码180个氨基酸,二级结构由8.9%的α–螺旋、47.8%的β–折叠和43.3%的无规则卷曲组成,其N端含有冷休克结构域。进化分析表明沙冬青与花生同源性最高。构建了AmCSDP真核表达载体Pcambia2300-35S-AmCSDP-OCS,经农杆菌介导转入本氏烟中。对转基因T1代和野生型烟草进行4℃低温胁迫处理,结果证实转基因本氏烟比野生型的耐冷能力强。 相似文献
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Expression profiling of transgenes (Cry1Ac and Cry2A) in cotton genotypes under different genetic backgrounds 下载免费PDF全文
Kashif NOOR Hafiza Masooma Naseer CHEEMA Asif Ali KHAN Rao Sohail Ahmad KHAN 《农业科学学报》2022,21(10):2818-2832
Transgenic cotton carrying the Cry1Ac gene has revolutionized insect pest control since its adoption, although the development of resistance in insect pests has reduced its efficacy. After 10 years of cultivating Bacillus thuringiensis (Bt) cotton with a single Cry1Ac gene, growers are on the verge of adopting Bt cotton that carries the double gene (Cry1Ac+Cry2A) due to its better effectiveness against insect pests. Thus, the current study was designed to evaluate the role of each gene in the effectiveness of Bt cotton carrying the double gene. The expression levels of the Cry1Ac and Cry2A genes were evaluated in the leaves of 10 genotypes (2 parents and 8 F1 hybrids) at 30 days after sowing (DAS), while samples of leaves, bolls and flowers were taken from the upper and lower canopies at 70 and 110 DAS. The F1 hybrids were developed through reciprocal crosses between two Bt (CKC-1, CKC-2) and two non-Bt (MNH-786, FH-942) parents. The differential expression of transgenes was evaluated through Enzyme Linked Immuno-Sorbent Assay (ELISA). The results showed that the MNH786×CKC-1 hybrid had the highest concentrations of Cry1Ac gene at 30 DAS (3.08 µg g–1) and 110 DAS (1.01 µg g–1) in leaves. In contrast, the CKC-2×MNH-786 hybrid showed the lowest concentrations of Cry1Ac gene at 30 DAS (2.30 µg g–1) and 110 DAS (0.86 µg g–1). The F1 hybrid FH-942×CKC-2 showed the highest concentrations of Cry2A gene at 30 DAS (8.39 µg g–1) and 110 DAS (7.74 µg g–1) in leaves, while the CKC-1×MNH-786 hybrid expressed the lowest concentrations of Cry2A gene at 30 DAS (7.10 µg g–1) and 110 DAS (8.31 µg g–1). A comparison between the two stages of plant growth showed that leaves had the highest concentrations at 30 DAS, whereas the lowest concentrations were observed at 110 DAS for both genes in leaves. When the expression pattern was compared between various plant parts in genotype CKC-2, it was found that leaves had higher concentrations of Cry1Ac (3.12 µg g–1) and Cry2A (8.31 µg g–1) at 70 DAS, followed by bolls (Cry1Ac (1.66 µg g–1) and Cry2A (8.15 µg g–1)) and flowers (Cry1Ac (1.07 µg g–1) and Cry2A (7.99 µg g–1)). The genotype CKC-2 had higher concentrations of Cry1Ac (3.12 µg g–1) and Cry2A (8.31 µg g–1) in the upper canopy but less accumulation (2.66 µg g–1 of Cry1Ac, 8.09 µg g–1 of Cry2A) in the lower canopy at 70 DAS. Similarly, at 110 DAS, the expression levels of Cry1Ac and Cry2A in upper and lower canopy leaves were 1.52 and 7.92 µg g–1, and 0.99 and 7.54 µg g–1, respectively. Hence, the current study demonstrates that different genotypes showed variable expression for both of the Cry1Ac and Cry2A genes during plant growth due to different genetic backgrounds. The Cry2A gene had three-fold higher expression than Cry1Ac with significant differences in expression in different plant parts. The findings of this study will be helpful for breeding insect-resistant double-gene genotypes with better gene expression levels of Cry1Ac and Cry2A for sustainable cotton production worldwide. 相似文献
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用不同浓度的和5-氮胞苷溶液配合不同时间处理了发生的在转录水平的转基因cry1Ab沉默水稻。结果表明,5-氮胞苷处理能使8%-30%转基因水稻植株的转基因cry1Ab恢复表达活性,复活性基因表达的Cry1Ab蛋白含量可达0.147%,45mg/L5-氮胞苷处理1d可使转基因cry1Ab的复活率及Cry1Ab蛋白含量达最高。同时,5-氮胞苷处理使转基因水稻植株出现矮化,分蘖数、单株粒重下降,穗长减小及开花期延迟等发育异常现象。5-氮胞苷作为一种弱诱变剂在克服转基因沉默及创造具优良农艺性状的转基因植物中具有一定的应用价值。 相似文献
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