本研究旨在提高黄瓜绿斑驳花叶病毒(cucumber green mottle mosaic virus,CGMMV)介导的基因沉默(VIGS)在瓜类作物中应用效率。以黄瓜、甜瓜和西瓜优良种质为实验材料,采用不同的CGMMV病毒载体接种方式、并设置不同的培养温度和相对湿度...本研究旨在提高黄瓜绿斑驳花叶病毒(cucumber green mottle mosaic virus,CGMMV)介导的基因沉默(VIGS)在瓜类作物中应用效率。以黄瓜、甜瓜和西瓜优良种质为实验材料,采用不同的CGMMV病毒载体接种方式、并设置不同的培养温度和相对湿度,以测试基因沉默的有效性。结果表明,真空渗透和种子吸胀的接种方式能够在瓜类作物中产生最高的基因沉默率(FGS)。在22℃和25℃的培养环境下,黄瓜、甜瓜和西瓜PDS沉默有效性(EGSL)较高。黄瓜、甜瓜和西瓜生长1个月后,EGSL达到100%,显著高于30℃的培养环境下EGSL。生长3个月后,甜瓜和西瓜在22℃培养环境下EGSL分别为89.7%和95%;在25℃培养环境下EGSL分别为85.6%和86.1%,均显著性高于30℃下EGSL。在相对湿度为30%和50%环境下,黄瓜的EGSL分别为70%和72%,甜瓜的EGSL分别为76%和73%,显著高于相对湿度为80%的培养环境下EGSL。西瓜在相对湿度为30%的培养环境下的EGSL为69%,显著高于相对湿度为50%时的EGSL(38%)和相对湿度在80%时的EGSL(33%)。综上所述,通过优化了瓜类作物的CGMMV-VIGS的技术体系中接种方式和培养环境参数,提高了基因沉默率和有效性,并且能够快速获得整个植株基因沉默的种质资源,为作物优质和抗逆基因功能的研究提供有效途径。展开更多
Activity of bc1 complex kinase(ABC1K)is an atypical protein kinase(aPK)that plays a crucial role in plant mitochondrial and plastid stress responses,but little is known about the responses of ABC1Ks to stress in cotto...Activity of bc1 complex kinase(ABC1K)is an atypical protein kinase(aPK)that plays a crucial role in plant mitochondrial and plastid stress responses,but little is known about the responses of ABC1Ks to stress in cotton(Gossypium spp.).Here,we identified 40 ABC1Ks in upland cotton(Gossypium hirsutum L.)and found that the Gh ABC1Ks were unevenly distributed across 17 chromosomes.The GhABC1K family members included 35 paralogous gene pairs and were expanded by segmental duplication.The GhABC1K promoter sequences contained diverse cis-acting regulatory elements relevant to hormone or stress responses.The qRT-PCR results revealed that most Gh ABC1Ks were upregulated by exposure to different stresses.Gh ABC1K2-A05 and Gh ABC1K12-A07 expression levels were upregulated by at least three stress treatments.These genes were further functionally characterized by virus-induced gene silencing(VIGS).Compared with the controls,the Gh ABC1K2-A05-and Gh ABC1K12-A07-silenced cotton lines exhibited higher malondialdehyde(MDA)contents,lower catalase(CAT),peroxidase(POD)and superoxide dismutase(SOD)activities and reduced chlorophyll and soluble sugar contents under NaCl and PEG stress.In addition,the expression levels of six stress marker genes(Gh DREB2A,Gh SOS1,Gh CIPK6,Gh SOS2,Gh WRKY33,and Gh RD29A)were significantly downregulated after stress in the Gh ABC1K2-A05-and Gh ABC1K12-A07-silenced lines.The results indicate that knockdown of Gh ABC1K2-A05 and Gh ABC1K12-A07 make cotton more sensitive to salt and PEG stress.These findings can provide valuable information for intensive studies of Gh ABC1Ks in the responses and resistance of cotton to abiotic stresses.展开更多
文摘本研究旨在提高黄瓜绿斑驳花叶病毒(cucumber green mottle mosaic virus,CGMMV)介导的基因沉默(VIGS)在瓜类作物中应用效率。以黄瓜、甜瓜和西瓜优良种质为实验材料,采用不同的CGMMV病毒载体接种方式、并设置不同的培养温度和相对湿度,以测试基因沉默的有效性。结果表明,真空渗透和种子吸胀的接种方式能够在瓜类作物中产生最高的基因沉默率(FGS)。在22℃和25℃的培养环境下,黄瓜、甜瓜和西瓜PDS沉默有效性(EGSL)较高。黄瓜、甜瓜和西瓜生长1个月后,EGSL达到100%,显著高于30℃的培养环境下EGSL。生长3个月后,甜瓜和西瓜在22℃培养环境下EGSL分别为89.7%和95%;在25℃培养环境下EGSL分别为85.6%和86.1%,均显著性高于30℃下EGSL。在相对湿度为30%和50%环境下,黄瓜的EGSL分别为70%和72%,甜瓜的EGSL分别为76%和73%,显著高于相对湿度为80%的培养环境下EGSL。西瓜在相对湿度为30%的培养环境下的EGSL为69%,显著高于相对湿度为50%时的EGSL(38%)和相对湿度在80%时的EGSL(33%)。综上所述,通过优化了瓜类作物的CGMMV-VIGS的技术体系中接种方式和培养环境参数,提高了基因沉默率和有效性,并且能够快速获得整个植株基因沉默的种质资源,为作物优质和抗逆基因功能的研究提供有效途径。
基金supported by the State Key Laboratory of Aridland Crop Science,Gansu Agricultural University,China(GSCS-2019-10)the National Natural Science Foundation of China(31801414 and 32260478)+2 种基金the Gansu Province Science and Technology Program,China(20JR10RA531)the Natural Science Foundation of Xinjiang Uygur Autonomous Region,China(2022D01E103)the Education Technology Innovation Project of Gansu Province,China(2022QB-076)。
文摘Activity of bc1 complex kinase(ABC1K)is an atypical protein kinase(aPK)that plays a crucial role in plant mitochondrial and plastid stress responses,but little is known about the responses of ABC1Ks to stress in cotton(Gossypium spp.).Here,we identified 40 ABC1Ks in upland cotton(Gossypium hirsutum L.)and found that the Gh ABC1Ks were unevenly distributed across 17 chromosomes.The GhABC1K family members included 35 paralogous gene pairs and were expanded by segmental duplication.The GhABC1K promoter sequences contained diverse cis-acting regulatory elements relevant to hormone or stress responses.The qRT-PCR results revealed that most Gh ABC1Ks were upregulated by exposure to different stresses.Gh ABC1K2-A05 and Gh ABC1K12-A07 expression levels were upregulated by at least three stress treatments.These genes were further functionally characterized by virus-induced gene silencing(VIGS).Compared with the controls,the Gh ABC1K2-A05-and Gh ABC1K12-A07-silenced cotton lines exhibited higher malondialdehyde(MDA)contents,lower catalase(CAT),peroxidase(POD)and superoxide dismutase(SOD)activities and reduced chlorophyll and soluble sugar contents under NaCl and PEG stress.In addition,the expression levels of six stress marker genes(Gh DREB2A,Gh SOS1,Gh CIPK6,Gh SOS2,Gh WRKY33,and Gh RD29A)were significantly downregulated after stress in the Gh ABC1K2-A05-and Gh ABC1K12-A07-silenced lines.The results indicate that knockdown of Gh ABC1K2-A05 and Gh ABC1K12-A07 make cotton more sensitive to salt and PEG stress.These findings can provide valuable information for intensive studies of Gh ABC1Ks in the responses and resistance of cotton to abiotic stresses.
