黍子NF-Y基因家族的鉴定及PmNF-YA8的功能分析OA
Identification of the NF-Y Gene Family and Functional Analysis of PmNF-YA8 in Broomcorn Millet
[目的]核因子 Y(nuclear factor Y,NF-Y)转录因子通过特异性结合靶基因启动子区 CCAAT 元件调控表达,在植物应对干旱、盐碱等非生物胁迫中起作用.黍子抗旱耐瘠,是盐碱地、边际土地不可或缺的作物,解析黍子 NF-Y基因家族功能,为丰富作物耐逆遗传理论、推动抗旱育种实践提供依据.[方法]利用黍子基因组数据和生物信息学方法,鉴定 NF-Y 家族成员,分析理化性质,构建系统进化树,分析保守结构域、基因结构和启动子顺式作用元件.用实时荧光定量 PCR 鉴定基因在不同部位的表达,并通过拟南芥转基因功能验证 PmNF-YA8在干旱胁迫下的表达特征.[结果]基于黍子参考基因组鉴定出 33 个 PmNF-Y 基因,包括 11 个 PmNF-YA、12 个 PmNF-YB 和 10 个 PmNF-YC 亚家族成员,蛋白长度为 122-571 个氨基酸,等电点为 4.74-10.19,分子量为 13.58-59.17 kDa.染色体定位显示,33 个基因(PmNF-YA1—PmNF-YA11、PmNF-YB1—PmNF-YB12和 PmNF-YC1—PmNF-YC10)分布于 15 条染色体.在 PmNF-Y 家族成员的启动子区域预测到 2 种干旱胁迫响应元件.黍子与 3 种植物(拟南芥、水稻和玉米)的 NF-Y 直系同源基因对分别为3、47 和 59 个.RT-qPCR 分析表明,PmNF-Y 基因在根、茎、叶等组织中广泛表达.干旱胁迫后,PmNF-YA8在小红黍和宁糜 9 号中均上调表达.经 300 mmol·L-1 甘露醇胁迫后,过表达 PmNF-YA8株系平均根长(14.03 mm)大于野生型拟南芥(9.07 mm),且 MDA 增幅(55.23%—57.12%)低于野生型(100.78%),SOD、POD 活性分别增加 24.28%—27.43%和 163.57%—341.33%,Pro 含量(151.31-175.14 μg·g-1)高于野生型(143.78 μg·g-1).[结论]在黍子中鉴定出 33 个 NF-Y 家族成员,从黍子中克隆了 PmNF-YA8,异源表达 PmNF-YA8通过调节胁迫相关基因的表达,增强了拟南芥植株对干旱胁迫的耐受性,可作为调控黍子抗旱性的重要候选基因.
[Objective]Nuclear factor Y(NF-Y)transcription factors regulate the expression of target genes by specifically binding to the CCAAT element in the promoter region of target genes,and play a role in plant responses to abiotic stresses such as drought and salinity.Broomcorn millet exhibits strong tolerance to drought and barrenness,serving as an indispensable crop for saline-alkali and marginal lands.Systematic elucidation of the functions of the NF-Y gene family in broomcorn millet will provide a theoretical basis for enriching the genetic theory of crop stress tolerance and promoting drought-resistant breeding practices.[Method]Genome data of broomcorn millet and bioinformatics approaches were employed to identify NF-Y family members,physicochemical properties,gene structures,construct phylogenetic trees,and predict conserved domains as well as cis-acting elements in promoters.Reverse Transcription Quantitative PCR(RT-qPCR)was used to determine the expression of these genes in different plant tissues,and the expression characteristics of PmNF-YA8 under drought stress were verified through transgenic functional validation in Arabidopsis thaliana.[Result]Based on the proso millet reference genome,33 PmNF-Y genes(including 11 PmNF-YA,12 PmNF-YB,and 10 PmNF-YC)were identified,encoding proteins with lengths of 122-571 amino acids,isoelectric points of 4.74-10.19,and molecular weights of 13.58-59.17 kDa.Chromosomal localization analysis showed that these 33 genes(PmNF-YA1-PmNF-YA11,PmNF-YB1-PmNF-YB12,and PmNF-YC1-PmNF-YC10)were distributed across 15 chromosomes.Two types of drought stress-responsive elements were predicted in the promoter regions of PmNF-Y family members.A total of 3,47,and 59 NF-Y orthologous gene pairs were identified between broomcorn millet and three other plant species(Arabidopsis thaliana,rice,and maize),respectively.RT-qPCR analysis revealed that PmNF-Y genes were widely expressed in tissues such as roots,stems,and leaves.The expression level of PmNF-YA8 was up-regulated in both XHS and NM9 varieties after drought stress.Transgenic Arabidopsis thaliana lines overexpressing PmNF-YA8 were generated,and phenotypic and physiological analyses showed that under 300 mmol·L-1 mannitol stress,the average root length of transgenic plants(14.03 mm)was longer than that of wild-type(WT)plants(9.07 mm).Additionally,transgenic plants had a lower MDA increase range(55.23%-57.12%)compared with WT plants(100.78%),with SOD and POD activities increased by 24.28%-27.43%and 163.57%-341.33%,respectively,and Pro content(151.31-175.14 μg·g-1)higher than that of WT plants(143.78 μg·g-1).[Conclusion]A total of 33 NF-Y family members were identified in broomcorn millet.The PmNF-YA8 gene was cloned from broomcorn millet and heterologously transformed into Arabidopsis thaliana.Its heterologous expression enhanced the drought stress tolerance of Arabidopsis thaliana plants by regulating the expression of stress-related genes,thus making PmNF-YA8 a key candidate gene for modulating drought tolerance in broomcorn millet.
宋裕祯;Bheel Chander Kumar;张颖星;王跃;Santra Dipak Kumar;曹晓宁;王瑞云
山西农业大学农学院,中国山西太谷 030801山西农业大学农学院,中国山西太谷 030801山西农业大学农学院,中国山西太谷 030801山西农业大学农学院,中国山西太谷 030801内布拉斯加大学林肯分校农艺系小宗粮豆研究与推广中心,美国内布拉斯加州斯克茨布拉夫 69361山西农业大学农学院,中国山西太谷 030801||山西农业大学农业基因资源研究中心,中国 太原 030031山西农业大学农学院,中国山西太谷 030801
黍子NF-YPmNF-YA8表达模式干旱胁迫
broomcorn milletNF-YPmNF-YA8expression patterndrought stress
《中国农业科学》 2026 (10)
2088-2108,21
现代农业产业技术体系建设专项(CARS-06-15.5-A16)、山西省现代农业产业技术体系建设(杂粮)专项资金(2026CYJSTX03-12)、山西省重点研发项目(2022ZDYF110)
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