基于转录组测序筛选新疆野苹果种子休眠解除过程中植物激素相关基因  

Screening of plant hormone-associated genes during seed dormancy release in Malus sieversii based on transcriptome sequencing

作　　者：房震 李静[1] 马娟[1] 张凯[1] 叶春秀 FANG Zhen;LI Jing;MAJuan;ZHANG Kai;YE Chunxiu(College of Forestry and Landscape Architecture,Xinjiang Agricultural University,Urumqi 830052,Xinjiang,China)

机构地区：[1]新疆农业大学林学与风景园林学院,乌鲁木齐830052

出　　处：《果树学报》2024年第10期1961-1978,共18页Journal of Fruit Science

基　　金：新疆维吾尔自治区自然科学基金面上项目(2023D01A28);新疆农业大学高层次人才引进项目(2521GCCRC)。

摘　　要：【目的】研究新疆野苹果(Malus sieversii)种子不同贮藏阶段转录组差异和激素含量变化,筛选新疆野苹果种子休眠解除过程中植物激素相关基因,为后续新疆野苹果种子休眠解除激素调控机制研究提供依据。【方法】以新疆野苹果种子为材料,对照未层积种子和经4℃低温沙藏层积处理(30、60、90、120 d)后种子进行转录组测序,采用酶联免疫吸附法测定种子中的脱落酸(abscisic acid,ABA)、赤霉素(gibberellin,GA)、生长素(auxin,IAA)、细胞分裂素(cytokinin,CTK)含量以及乙烯氨基环丙烷羧酸氧化酶(1-aminocyclopropane-1-carboxylic acid oxidase,ACO)和氨基环丙烷羧酸合成酶(1-aminocyclopropane-1-carboxylic acid oxidase synthase,ACS)的活性。【结果】新疆野苹果种子GA、IAA、CTK含量随贮藏时间的增加呈上升趋势,而ABA含量呈下降趋势。乙烯(ethylene,ETH)合成途径ACO和ACS活性随贮藏时间的增加呈增强趋势。GO富集分析筛选出85个差异基因调控种子萌发(GO:0010029),114个GA相关差异基因,313个ABA相关差异基因和156个ETH相关差异基因。KEGG通路富集分析显示,主要富集通路有植物激素信号转导、MAPK信号通路-植物、内质网中的蛋白加工、淀粉和蔗糖代谢、糖酵解/糖原生成等。ABA信号传导通路中有3个PYR/PYL基因下调表达,2个蛋白磷酸酶2C(PP2C-type protein phosphatases,PP2C)、3个蔗糖非发酵相关的蛋白激酶(sucrose non-fermenting-1-related protein kinase 2,SnRK2)上调表达和2个ABA分解代谢8'-羟化酶(8'-hydroxylases)基因表达显著上调。GA信号转导途径中2个受体GID1(gibberellin insensitive dwarf 1)和6个负调控因子DELLA蛋白上调表达;ETH受体ETR(Ethylene receptor)的直接前体1-氨基环丙烷-1-羧酸(1-aminocyclopropane-1-carboxylic acid,ACC)和ACO基因上调表达。【结论】新疆野苹果种子经低温沙藏层积处理,ERF2-like表达量与ACO、ACS活性变化呈相反趋势,PYR1-like、WRKY33基因表达�【Objective】The study aimed to study the transcriptome differences and hormone content changes of the seeds of Malus sieversii at different stages of stratification in order to screen the plant hormones genes related to dormancy release and provide a basis for subsequent studies on the hormonal regulation mechanism of seed dormancy release in M.sieversii.【Methods】The seeds of M.sieversii were used as materials,transcriptome sequencing was performed on the seeds at different stages of the stratification at 4℃(0,30,60,90 and 120 d).The content of abscisic acid(ABA),gibberellins(GA),auxin(IAA),cytokinin(CTK),the activities of ethylene 1-aminocyclopropane-1-carboxylate oxidase(ACO)and 1-aminocyclopropane-1-carboxylate oxidase synthase(ACS)were measured using an enzyme-linked immunosorbent assay.【Results】The ABA content of the seeds showed a decreasing trend with the increase of stratification time,the maximum content was 80.22 ng·g-1on 0 d of the storage,the content on 60 d of the storage was significantly lower than that at the three periods of 0,30 and 90 d(p<0.05),the content reached a minimum of 43.67 ng·g-1on 120 d.The GA content showed an increasing trend with the increase of the storage time,the content was the lowest at 78.42 pmol·g-1on 0 d,and the content on 120 d significantly higher than those of the other periods(p<0.05),and reached a maximum of 170.67 pmol·g-1.The IAA content showed an increasing trend with the storage time,and the content on 90 d was significantly higher than those of the other periods,reaching a maximum value of41.36 nmol·g-1.The CTK content on 90 d was significantly higher than those of the other periods,reaching a minimum value of 43.67 ng·g-1,it showed a decreasing trend in the storage time of 0-30 d,the content on 30 d was significantly lower than those of the other periods(p<0.05),reaching a minimum value of 31.34 ng·g-1,and showed an increasing trend in the storage time of 30-60 d,suggesting that CTK would promote the accumulation of seed assimilates in this p

关 键 词：新疆野苹果 转录组 内源激素 差异表达基因 种子休眠 

分 类 号：S661.1[农业科学—果树学]