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乌菜BcVIL2基因克隆及春化响应表达分析 被引量:4
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作者 桂尚枝 刘雪晴 +7 位作者 王英 唐小燕 赵龙龙 李广 吴思文 温宏伟 汪承刚 陈国户 《西南农业学报》 CSCD 北大核心 2023年第9期1843-1851,共9页
【目的】从乌菜(Brassica campestris L.)中克隆VIN3⁃LIKE 2(VIL2)基因并进行生物信息学与表达分析,为进一步研究BcVIL2基因功能奠定基础。【方法】以‘徽乌19’为试验材料,利用同源克隆等技术获得BcVIL2基因,对其序列进行蛋白分子量(MW... 【目的】从乌菜(Brassica campestris L.)中克隆VIN3⁃LIKE 2(VIL2)基因并进行生物信息学与表达分析,为进一步研究BcVIL2基因功能奠定基础。【方法】以‘徽乌19’为试验材料,利用同源克隆等技术获得BcVIL2基因,对其序列进行蛋白分子量(MW)、糖基化位点、二级结构等生物信息学分析;利用荧光定量PCR等技术分析其应答春化表达模式。【结果】BcVIL2基因ORF(Open reading fragment)全长为1296 bp,共编码431个氨基酸,属于亲水性蛋白,预测含有1个潜在的糖基化位点和59个潜在的磷酸化位点,蛋白分子量为47.91 kDa,理论等电点(pI)为8.27;其二级结构主要由α⁃螺旋与无规则卷曲组成;亚细胞定位预测其可能在细胞核中发挥作用;同源序列比对分析显示该基因与芸薹属植物VIL2基因同源性较高;亲缘进化树分析也显示该基因与白菜(B.rapa,syn.B.campestris)和芥菜(B.juncea)VIL2基因亲缘关系很近;荧光定量PCR结果表明,盛花期时BcVIL2基因在根中表达量最低,在荚中表达量最高,而叶中表达量显著低于茎和花中表达量;一月苗龄植株春化后BcVIL2基因的表达量显著低于未春化时的表达量。【结论】乌菜VIL2基因响应春化负表达,与春化反应相关基因BraA01g032910.3C、BraA05g026410.3C与BraA10g016160.3C密切相关,可能存在互作关系,本研究结果为深入分析BcVIL2基因的功能提供了良好基础。 展开更多
关键词 乌菜 vil2 春化 基因克隆 表达分析
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Intron-retained alternatively spliced VIN3-LIKE 2 gene from Chimonanthus praecox promotes flowering in transgenic Arabidopsis 被引量:2
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作者 Tian Xu Ning Liu +4 位作者 Chunyu Wu Yi Zhang Mingkang Tian Shunzhao Sui Zhineng Li 《Horticultural Plant Journal》 SCIE CAS CSCD 2023年第4期789-800,共12页
Vernalization is a process of acquiring or accelerating the flowering ability by prolonged cold exposure.VERNALIZATION INSENSITIVE3(VIN3)is induced by chilling and is extremely important for the vernalization response... Vernalization is a process of acquiring or accelerating the flowering ability by prolonged cold exposure.VERNALIZATION INSENSITIVE3(VIN3)is induced by chilling and is extremely important for the vernalization response in Arabidopsis thaliana.However,the issue of the induction of the VIN3-LIKE genes in wintersweet(Chimonanthus praecox)has been largely neglected.In the present study,we explored the molecular regulation of the PHD type finger protein-encoding gene CpVIL2 in relation to the growth and development of wintersweet in Arabidopsis.In wintersweet,quantitative real-time PCR(qRT-PCR)analysis showed that the relative expression of CpVIL2-As2i(intron-retained alternatively spliced in the second intron)was extremely higher in the pistils than in the other tissues.And the relative CpVIL2-As2i expression in flower buds(FBs)treated at 8°C was higher than that of FBs in December,2016 under natural conditions,which was not detected in non-flowering FBs at 16°C.In Arabidopsis,the expression patterns of the CpVIL2-As2i gene were detected at first in CpVIL2-As2i pro::GUS(β-glucuronidase)lines,with predominantly higher expression in flowers and inflorescence.Meanwhile,the hormone-induced expression profiles of the CpVIL2-As2i promoter were confirmed using exogenous induction by abscisic acid(ABA)and indole acetic acid(IAA)phytohormones,where the GUS enzyme activity obviously decreased compared with that of control.In comparison with Arabidopsis/Col-0,early flowering was detected in ectopic 35S::CpVIL2-As2i lines.Overall,these results demonstrated the function of the CpVIL2-As2i gene,at the same time,provided us with new insights into the molecular mechanisms of early flowering and complex regulatory networks of vernalization in wintersweet. 展开更多
关键词 Chimonanthus praecox PHD finger protein vil2 Ectopic expression Flowering time VERNALIZATION
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