油茶NAC基因鉴定及对干旱胁迫响应分析
Identification of NAC Gene in Camellia oleifera and Analysis of its response to drought stress
[目的]NAC是参与调控植物逆境胁迫反应的的一类特异转录因子,在植物应对干旱胁迫中具有重要的调控作用。油茶是南方丘陵山地重要的经济树种,主产区夏季干旱严重影响了油茶生产。研究油茶应对干旱胁迫的响应机制具有重要意义。[方法]为明确油茶NAC基因家族成员并探究其对干旱胁迫响应的表达特征,本研究以油茶全长转录组为参考,通过蛋白序列比对和保守功能域,鉴定了 67个CoNAC 基因家族成员,进一步明确了各基因的结构、理化性质、系统进化关系、亚细胞定位,基于油茶干旱转录组数据,明确各CoNAC在油茶不同干旱程度和复水下的表达水平,并采用qPCR对CoNAC在不同油茶品种和不同干旱胁迫程度下的表达特征进行验证。[结果]根据系统发育关系,67 个 CoNAC 可分为 17 个亚类,ANAC2 亚类最多,有 13个成员。亚细胞定位预测表明,大部分 CoNAC 蛋白都定位在细胞核,少数定位在细胞质、线粒体、叶绿体或细胞外。基于不同干旱程度和复水的干旱转录组数据,CoNAC 基因间的表达模式存在一定差异,且发现在中度干旱、重度干旱胁迫时期,CoNAC28、CoNAC51、CoNAC52、CoNAC56、CoNAC60 和 CoNAC61 等基因随着干旱胁迫程度加强,其表达量持续增加。在复水后,CoNAC28、CoNAC51、CoNAC52、CoNAC56、CoNAC60、和CoNAC61等基因均无表达或表达呈明显下降趋势。qPCR验证结果显示,长林53油茶品种中,6 个 CoNAC 基因在干旱处理时表达量均呈显著上调趋势,其表达量都表现出先上升后下降的趋势。长林18 油茶中,CoNAC28、CoNAC56、CoNAC60 和 CoNAC61 基因在干旱胁迫下的表达量均呈显著上调趋势。[结论]CoNAC 基因表达受不同程度干旱胁迫的调控,且其表达模式和表达水平在不同油茶品种间存在差异,表明油茶不同品种的耐旱性不同。本研究为油茶 NAC 转录因子进一步功能分析奠定了理论基础,为油茶耐旱性的遗传改良提供参考依据。
[Objective]NAC is a kind of specific transcription factors which is involved in the regulation of plant stress response.It plays an important role in the regulation of plants in response to drought stress.Camellia oleifera is an important economic tree species in southern hills and mountains.the summer drought in the main producing areas has seriously affected the production of C.oleifera.It is of great significance to study the response mechanism of C.oleifera to drought stress.[Method]In order to explore the expression characteristics of NAC gene family members of C.oleifera to resistance drought stress,the full-length transcriptome of C.oleifera was used as a reference,through protein sequence alignment and conserved functional domain.67 members of CoNAC gene family were identified,and the structure,physical and chemical properties,phylogenetic relationship and subcellular location of each gene were further identified.Based on the data of Camellia oleifera drought transcriptome,the expression levels of CoNAC in C.oleifera under different drought degrees and rewatering were determined.And qPCR was used to verify the expression characteristics of CoNAC in different C.oleifera varieties and different degrees of drought stress.[Result]According to the phylogenetic relationships,67 NAC genes were divided into 17 subfamilies,of which ANAC2 subclass with 13 members is the most.The prediction of subcellular localization showed that most of the CoNAC proteins were located in the Nucleus and a few were located in the cytoplasm,mitochondria,chloroplasts or extracellular.Based on the drought transcriptome data of different drought degrees and rewatering,there were some differences in the expression patterns of most CoNAC genes.It was also found that during the period of moderate and severe drought stress,the genes CoNAC28,CoNAC51,CoNAC52,CoNAC56,CoNAC60 and CoNAC61 increased with the degree of drought stress,Its expression continued to increase.After rehydration,CoNAC28,CoNAC51,CoNAC52,CoNAC56,CoNAC60 and CoNAC61 genes were not expressed or decreased significantly.qPCR verification results show,In'Changlin 53',the expression of six CoNAC genes was significantly up-regulated under drought treatment,and the expression increased at first and then decreased.In'Changlin 18',the expressions of CoNAC28,CoNAC56,CoNAC60 and CoNAC61 genes were significantly up-regulated under drought stress.[Conclusion]the results show that the expression of CoNAC gene was regulated by different level of drought stress,and its expression pattern varied among different C.oleifera varieties,which indicated different C.oleifera varieties have different drought tolerance.This study laid a theoretical foundation for further functional analysis of NAC transcription factors in C.oleifera and provided a reference basis for genetic improvement of drought tolerance in C.oleifera.
曹瑞兰;李知青;欧阳雯婷;胡冬南;周增亮;苏文娟;陈霞;刘娟;
江西农业大学林学院江西省森林培育重点实验室,南昌 330045;江西农业大学林学院江西省森林培育重点实验室,南昌 330045;福建农林大学林学院,福建 350028;江西农业大学林学院江西省森林培育重点实验室,南昌 330045;江西农业大学林学院江西省森林培育重点实验室,南昌 330045;江西农业大学林学院江西省森林培育重点实验室,南昌 330045;江西农业大学林学院江西省森林培育重点实验室,南昌 330045;江西农业大学林学院江西省森林培育重点实验室,南昌 330045;
Camellia oleifera NAC gene family bioinformatics analysis drought stress
1065-1080 / 16
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