油茶PODs基因鉴定及在自交和异交下的表达分析
作者:
作者单位:

1.中南林业科技大学林学院经济林培育与保护教育部重点实验室,长沙410004;2.湖南省林业科学院油茶研究所,长沙410004

作者简介:

研究方向为经济林栽培育种,E-mail: 1873079745@qq.com

通讯作者:

周俊琴,研究方向为经济林栽培育种,E-mail:zhoujunqin@csuft.edu.cn

中图分类号:

基金项目:

湖南省林业科技攻关与创新项目(XLKY202323,XLK201939);中南林业科技大学研究生科技创新项目(2023CX02043)


Expression Analysis under Self and Outcross Pollination and Identification of PODs Gene in Camellia oleifera Abel.
Author:
Affiliation:

1.Key Laboratory of Cultivation and Protection for Non-wood Forest Trees, Ministry of Education, College of Forestry, Central South University of Forestry and Technology, Changsha 410004;2.Camellia oleifera Abel. Research Institute, Hunan Academy of Forestry, Changsha 410004

Fund Project:

Forestry Science and Technology Research and Innovation Project of Hunan Province (XLKY202323,XLK201939); Science Innovation Project for Post-graduates of Central South University of Forestry and Technology (2023CX02043)

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    摘要:

    油茶(Camellia oleifera Abel.)具有自交不亲和性(SI,self-incompatibility),自然座果率低,这严重影响油茶产量,制约了油茶产业的发展。为研究过氧化物酶(POD,peroxidase)在油茶自交不亲和反应中的作用,本研究通过逆转录克隆技术从油茶中克隆出4条POD基因,分别命名为CoPOD1/2/3/4。其基因编码区长度分别为1086、1011、1020和1218 bp,编码361、336、339和405个无跨膜结构、有信号肽的蛋白质。同源序列比对显示CoPOD1/2/3/4蛋白质之间的同源性较低,但均有过氧化物酶活性位点和过氧化酶近端血红素配体序列;系统进化树显示CoPOD1/2/3/4蛋白质与茶树(Camellia sinensis(L.)O. Ktze.)POD蛋白质的亲缘关系最近。实时荧光定量结果显示,油茶PODs基因在自交授粉后24~48 h范围内呈现显著上调后又下降的趋势,在自交授粉36 h雌蕊中CoPOD1/3/4的表达量高于异交。自交授粉24~72 h油茶雌蕊内POD酶活性高于异交并在36 h达到最高值,异交授粉雌蕊内POD活性前期处于动态平衡状态,在72 h后出现明显上调,据此推测油茶PODs基因可能参与了油茶自交授粉后花粉管程序性死亡,进而参与了油茶自交不亲和反应。本研究为进一步研究油茶自交不亲和机制提供参考。

    Abstract:

    Camellia oleifera Abel., which shows self-incompatibility (SI) and low natural fruit setting rate, has seriously affected its yield production and the following development in industry. To uncover the function of Peroxidase (POD) in self-incompatibility, four POD genes CoPOD1/2/3/4 were cloned from C. oleifera by reverse transcription cloning technology. The length of gene coding regions were 1086, 1011, 1020 and 1218 bp, encoding 361, 336, 339 and 405 amino acids non-transmembrane proteins with signaling peptides, respectively. Although lower sequence similarity among the CoPOD1/2/3/4 proteins is observed, they all contain peroxidase active sites and heme ligand sequences near peroxidase. The phylogenetic analysis revealed closely sequence homology of CoPOD1/2/3/4 to those of Camellia sinensis (L.) O. Ktze. Real-time fluorescence quantitative results showed that PODs were significantly up-regulated, followed by decreased expression within 24-48 h after self-pollination. The transcripts of CoPOD1/3/4 were higher than cross-pollinated in pistills at 36 h after self-pollination. The POD activity in the pistil of self-pollination was higher than that of cross-pollinated within 24-72 h, showing the highest expression peak at 36 h. The POD activity in the pistil of the cross-pollinated was in dynamic balance at early stage, with significantly up-regulated after 72 h. We speculate that the POD genes might be involved in the programmed pollen tube death of C. oleifera after self-pollination, thus participating in the self-incompatibility reaction. Collectively, this paper provides reference for further research on self-incompatibility mechanism in C. oleifera.

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姚亚轩,袁军,卢梦琪,等.油茶PODs基因鉴定及在自交和异交下的表达分析[J].植物遗传资源学报,2023,24(5):1472-1484.

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  • 收稿日期:2023-03-09
  • 最后修改日期:2023-04-10
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  • 在线发布日期: 2023-08-30
  • 出版日期: 2023-08-30
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