桃生长势相关基因PpSAUR73功能鉴定

doi:10.3864/j.issn.0578-1752.2023.20.012

中国农业科学 ›› 2023, Vol. 56 ›› Issue (20): 4072-4086.doi: 10.3864/j.issn.0578-1752.2023.20.012

桃生长势相关基因PpSAUR73功能鉴定

杨丽¹(), 曹洪波¹, 张学英¹, 翟含含², 李辛淼¹, 彭佳伟¹, 田义³(), 陈海江¹()

¹ 河北农业大学园艺学院，河北保定 071000
² 喀什职业技术学院，新疆喀什 844000
³ 河北农业大学山区研究所/河北省山区农业技术创新中心/ 国家北方山区农业工程技术研究中心，河北保定 071001

收稿日期:2023-03-24 接受日期:2023-06-30 出版日期:2023-10-16 发布日期:2023-10-31
通信作者:
陈海江，chenhaijiang2001@163.com。
田义，tianyi@hebau.edu.cn
联系方式: 杨丽，E-mail：1097125451@qq.com。
基金资助:
财政部和农业农村部：国家现代农业产业技术体系资助项目(CARS-30-2-03); 河北省重点研发计划(20326804D); 热杂果现代种业科技创新团队(21326310D)

Functional Identification of Peach Gene PpSAUR73

YANG Li¹(), CAO HongBo¹, ZHANG XueYing¹, ZHAI HanHan², LI XinMiao¹, PENG JiaWei¹, TIAN Yi³(), CHEN HaiJiang¹()

¹ Horticultural Department, Agricultural University of Hebei, Baoding 071000, Hebei
² Kashgar Vocational and Technical College, Kashgar 844000, Xinjiang
³ Mountainous Areas Research Institute, Hebei Agricultural University/Technology Innovation Center for Agriculture in Mountainous Areas of Hebei Province/National Engineering Research Center for Agriculture in Northern Mountainous Areas, Baoding 071001, Hebei

Received:2023-03-24 Accepted:2023-06-30 Published:2023-10-16 Online:2023-10-31

摘要/Abstract

摘要：

【目的】克隆桃树势相关基因PpSAUR73，分析其对多种激素的响应，鉴定其在调控拟南芥生长势中的作用，为科学合理调控树势提供分子依据。【方法】以‘中油蟠9号’为材料进行激素处理，利用实时荧光定量分析PpSAUR73在24 h内的动态响应;以桃品种‘久艳’为材料克隆PpSAUR73;构建PpSAUR73过量表达载体，将其转化拟南芥;对转基因拟南芥进行表型观察，对同时播种的转基因与野生型拟南芥进行萌芽率统计，对萌发一致的生长7 d拟南芥进行根长及下胚轴的测量，对萌发一致的拟南芥进行不同浓度的激素处理;取7 d大小的两个转基因株系及野生型拟南芥材料进行转录组测序，对差异表达基因进行功能分析、KEGG通路富集分析，并分析调控基因。【结果】PpSAUR73能够对激素处理做出快速响应。过表达PpSAUR73能够影响拟南芥种子萌芽，转基因拟南芥幼苗下胚轴及根长较野生型长，莲座大，整体长势好于野生型，且降低了对生长素的敏感性。过表达PpSAUR73的转录组分析结果显示，在两个对比组中均表达的差异基因有128个，其中有84个上调基因，44个下调基因，并对20个表达量较高的差异基因进行了描述。对过表达PpSAUR73产生的差异表达基因进行GO功能显著性富集分析，结果表明差异表达基因在细胞组分方面富集的基因最多，定位在细胞质、细胞膜、细胞器和细胞外区域。对差异表达基因进行KEGG通路富集分析，结果表明差异表达基因主要富集到苯丙氨酸生物合成通路、植物激素信号转导通路、淀粉蔗糖代谢通路等代谢通路。在苯丙氨酸生物合成通路中，PpSAUR73能够调控编码过氧化物酶基因AT1G05260、AT3G01190、AT3G32980、AT5G15180表达上调，过氧化物酶与木质素合成相关，木质素含量与植株长势呈显著相关，暗示过表达PpSAUR73可能参与调控拟南芥木质素合成从而调控长势。在植物激素信号转导通路中，生长素信号转导中的一些生长素响应基因AtSAUR41、AtSAUR71、AtSAUR51、AtSAUR72和AtSAUR1等表达下调，脱落酸信号转导途径中磷酸酶蛋白AtPP2CA表达上调，而脱落酸信号通路基因AtPYL5表达下调。PpSAUR73能够调控拟南芥长势，参与多种激素信号转导。【结论】PpSAUR73能够快速对激素做出响应，且能够调控转基因拟南芥长势;PpSAUR73过量表达导致的差异基因主要富集到苯丙氨酸生物合成通路、植物激素信号转导通路、淀粉蔗糖代谢通路等代谢通路;PpSAUR73调控IAA、ABA信号转导，推测其在桃树的生长发育过程中起到了重要作用。

关键词: 桃, PpSAUR73, 基因表达, 激素处理, 信号转导

Abstract:

【Objective】 The object of this study was to isolate a peach potential-related gene PpSAUR73, to analyze its expression response to hormones, and to identify its role in regulating seedling growth in transgenic Arabidopsis, so as to provide the molecular basis for the regulation of tree potential. 【Method】 Using Zhongyou Pan 9 as the material for hormone treatment, the real-time fluorescence quantitative analysis was used to analyze the dynamic response of PpSAUR73 within 24 hours. PpSAUR73 was cloned from the peach variety Jiuyan. PpSAUR73 overexpression vector was constructed and transformed into Arabidopsis. Phenotypic observation of genetical modified Arabidopsis was carried out, and the germination rate statistics of both genetically modified and wild-type Arabidopsis sown simultaneously were performed too. The root length and hypocotyl of 7-day growing Arabidopsis with consistent germination were measured, and Arabidopsis with consistent germination was treated with different concentration hormone. Transcriptome sequencing was performed using 7-day-old seedlings, and the differentially expressed genes were analyzed by functional analysis, KEGG pathway enrichment analysis, and regulatory genes analysis, respectively. 【Result】 PpSAUR73 could respond quickly to hormone treatments. The overexpression of PpSAUR73 could affect the germination of Arabidopsis seeds. The hypocotyl and root length of seedlings were longer than those of wild type. In addition, the rosette of transgenic Arabidopsis was larger, and the overall growth potential was larger than wild type. The transgenic Arabidopsis showed decreased sensitivity to auxin. The transcriptome analysis of overexpressing PpSAUR73 showed that there were 128 differentially expressed genes in both control groups, including 84 up-regulated genes and 44 down-regulated genes, and 20 differentially expressed genes were described. The GO function significant enrichment analysis of the differentially expressed genes generated by overexpression of PpSAUR73 showed that the differentially expressed genes were the most abundant in cell components, located in cytoplasm, cell membrane, organelle and extracellular regions. KEGG pathway enrichment analysis on differentially expressed genes were conducted, and the results showed that the differentially expressed genes in pairwise comparisons CK vs SAUR73-1 and CK vs SAUR73-14 were mainly enriched in phenylalanine biosynthesis pathway, plant hormone signal transduction pathway, starch sucrose metabolic pathway and other metabolic pathways. In the phenylalanine biosynthesis pathway, PpSAUR73 could regulate the upregulation of peroxidase encoding genes AT1G05260, AT3G01190, AT3G32980 and AT5G15180. Peroxidases were associated with lignin synthesis, and lignin content was significantly correlated with plant growth, suggesting that overexpression of PpSAUR73 might be involved in regulating lignin synthesis in Arabidopsis and thus growth. In plant hormone signal transduction pathway, the expression of some auxin responsive genes of AtSAUR41, AtSAUR71, AtSAUR51, AtSAUR72 and AtSAUR1 in auxin signal transduction pathway was down-regulated, the expression of phosphatase protein AtPP2CA in abscisic acid signal transduction pathway was up-regulated, and the expression of abscisic acid signal pathway gene AtPYL5 was down-regulated. PpSAUR73 could regulate the growth of Arabidopsis and participate in multiple hormone signal transduction pathways. 【Conclusion】 This study found that PpSAUR73 could quickly respond to hormones and regulate the growth in transgenic Arabidopsis. The differentially expressed genes caused by overexpressed genes caused by overexpression of PpSAUR73 were mainly enriched in metabolic pathways, such as phenylalanine biosynthesis pathway, plant hormone signaling pathway, and starch sucrose metabolism pathway. PpSAUR73 also played an important role in IAA and ABA signal transduction pathways, it was speculated that it played an important role in the growth and development of peach trees.

Key words: peach, PpSAUR73, gene expression, hormone treatment, signal transduction

杨丽, 曹洪波, 张学英, 翟含含, 李辛淼, 彭佳伟, 田义, 陈海江. 桃生长势相关基因PpSAUR73功能鉴定[J]. 中国农业科学, 2023, 56(20): 4072-4086.

YANG Li, CAO HongBo, ZHANG XueYing, ZHAI HanHan, LI XinMiao, PENG JiaWei, TIAN Yi, CHEN HaiJiang. Functional Identification of Peach Gene PpSAUR73[J]. Scientia Agricultura Sinica, 2023, 56(20): 4072-4086.

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链接本文: https://www.chinaagrisci.com/CN/10.3864/j.issn.0578-1752.2023.20.012

https://www.chinaagrisci.com/CN/Y2023/V56/I20/4072

图/表 13

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表2

SAUR73部分共同差异基因的信息描述"

基因ID Gene ID	描述 Description	KEGG通路 KEGG pathway
AT1G23720	未命名蛋白物质 Unnamed protein product
AT1G43790	气管元件分化相关6 Tracheary element differentiation-related 6
AT1G43800	植物硬脂酰酰基载体蛋白去饱和酶家族蛋白 Plant stearoyl-acyl-carrier-protein desaturase family protein	脂肪酸生物合成（ko00061），生物合成不饱和脂肪酸（ko01040），脂肪酸代谢（ko01212） Fatty acid biosynthesis (ko00061), Biosynthesis of unsaturated fatty acids (ko01040), Fatty acid metabolism (ko01212)
AT1G47395	假设蛋白质AT1G47395 Hypothetical protein AT1G47395
AT1G56600	半乳糖醇合成酶 Galactinol synthase 2	半乳糖代谢（ko00052） Galactose metabolism (ko00052)
AT1G60190	ARM重复超家族蛋白 ARM repeat superfamily protein	泛素介导的蛋白水解（ko04120） Ubiquitin mediated proteolysis (ko04120)
AT1G77120	乙醇脱氢酶1 Alcohol dehydrogenase 1	糖酵解/糖异生（ko00010），脂肪酸降解（ko00071），酪氨酸代谢（ko00350），亚麻酸代谢（ko00592） Glycolysis/Gluconeogenesis (ko00010), Fatty acid degradation (ko00071), Tyrosine metabolism (ko00350), alpha-Linolenic acid metabolism (ko00592)
AT3G12830	SAUR样生长素反应蛋白家族 SAUR-like auxin-responsive protein family	植物激素信号转导（ko04075） Plant hormone signal transduction (ko04075)
AT3G14440	9-顺式环氧类胡萝卜素双加氧酶 Nine-cis-epoxycarotenoid dioxygenase 3	类胡萝卜素生物合成（ko00906） Carotenoid biosynthesis (ko00906)
AT3G28550	富含脯氨酸的延伸蛋白家族蛋白 Proline-rich extensin-like family protein
AT3G43190	蔗糖合酶4 Sucrose synthase 4	淀粉和蔗糖代谢（ko00500） Starch and sucrose metabolism (ko00500)
AT4G33070	硫胺素焦磷酸依赖的丙酮酸脱羧酶家族蛋白 Thiamine pyrophosphate dependent pyruvate decarboxylase family protein	糖酵解/糖异生（ko00010） Glycolysis/Gluconeogenesis (ko00010)
AT5G04150	未知的，偏 Unknown, partial
AT5G35190	假设蛋白质AXX17_AT5G31360 Hypothetical protein AXX17_AT5G31360
ATCG00430	NADH脱氢酶亚基K NADH dehydrogenase subunit K	氧化磷酸化（ko00190） Oxidative phosphorylation (ko00190)
ATCG00470	ATP合成酶CF1 epsilon亚基 ATP synthase CF1 epsilon subunit	氧化磷酸化（ko00190），光合作用（ko00195） Oxidative phosphorylation (ko00190), Photosynthesis (ko00195)
ATCG00530	包膜蛋白 Envelope membrane protein	光合作用（ko00195） Photosynthesis (ko00195)
ATCG00670	ATP依赖性Clp蛋白酶蛋白水解亚基 ATP-dependent Clp protease proteolytic subunit
ATCG00810	核糖体蛋白L22 Ribosomal protein L22	核糖体（ko03010） Ribosome (ko03010)
ATCG01020	核糖体蛋白L32 Ribosomal protein L32	核糖体（ko03010） Ribosome (ko03010)

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基因 Gene	引物 Primer sequences (5'-3')
PpActin	F: GCCCAAGTGCTTCGTATGCT
PpActin	R: ATCACCGGCTGCAATCCA
PpSAUR73	F: GGCACCCAACTGAGAAGTGA
PpSAUR73	R: ACGAGGTTCCAAAAGGGCAT

基因ID Gene ID	Log₂FC （CK/SAUR73-1）	Log₂FC （CK/SAUR73-14）	描述 Description
AT1G05260	1.20742752491825	1.00323218173246	过氧化物酶超家族蛋白 Peroxidase superfamily protein
AT1G26390	-2.70470526776539	-3.79852645503335	FAD结合黄连素家族蛋白 FAD-binding Berberine family protein
AT1G66270	1.56761594119799	1.4138309067234	糖基水解酶超家族蛋白 Glycosyl hydrolase superfamily protein
AT1G66280	1.32994741757618	1.28961957063889	糖基水解酶超家族蛋白 Glycosyl hydrolase superfamily protein
AT1G66800	1.24780969170763	1.32419227545878	NAD(P)结合罗斯曼折叠超家族蛋白NAD(P)-binding Rossmann-fold superfamily protein
AT2G32860	1.23824493211364	1.37258943978524	β-葡萄糖苷酶33 Beta glucosidase 33
AT3G01190	1.34813864857128	1.07544938464363	过氧化物酶超家族蛋白 Peroxidase superfamily protein
AT3G32980	1.38165803707019	1.11752278178902	过氧化物酶超家族蛋白 Peroxidase superfamily protein
AT3G49110	1.28803391956951	1.78265014560853	过氧化物酶CA peroxidase CA
AT5G15180	1.34289458873555	1.22134563236366	过氧化物酶超家族蛋白 Peroxidase superfamily protein
AT5G28510	1.77489118882188	1.59816016112596	β-葡萄糖苷酶24 Beta glucosidase 24

桃生长势相关基因PpSAUR73功能鉴定

Functional Identification of Peach Gene PpSAUR73

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