CaCl2处理对‘伏脆蜜’枣裂果生理特性及基因表达与代谢物的影响.pdf
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1、分类号:S665.1 密级:公开单位代码:10757学 号:10757193015珞里木士当TARIM UNIVERSITY硕士学位论文CaCb处理对伏脆蜜枣裂果生理特性及 基因表达与代谢物的影响Effects of CaCh treatment on physiological characteristics,gene expression and metabolites of fucuimi Jujube fruit cracking研究生姓名:张晶晶指导教师:林敏娟 教授申请学位门类级别:农学硕士专业名称:果树学研究方向:果树种质资源与遗传育种所在学院:园艺与林学学院新疆-阿拉尔 二。二
2、二年六月(C)1994-2023 China Academic Journal Electronic Publishing House.All rights reserved,http:/学位论文独创性声明本人郑重声明:所呈交的学位论文是本人在导师指导下进行的研究工作及 取得的研窕成果。据我所知,除了文中特别加以标注和致谢的地方外,论文中 不包含其他个人或集体已经发表或撰写过的研究成果。对本人研究做出重要贡 献的个人及集体,均已在论文中作了明确的说明并表示谢意。本人完全意识到 本声明的法律结果由本人承担。学位论文作者签名:注晶晶签字日期:入4年/月牛日学位论文版权认定和使用授权书本人完全了解学
3、校有关保留、使用学位论文的规定,有权保留并向国家有 关部门或机构送交论文的复印件和磁盘,允许论文被查阅和借用。本人授权塔 里木大学可以将学位论文的全部或部分内容编入有关数据库进行检索,可以采 用影印、缩印或扫描等复制手段保存、汇编学位论文。(保密的学位论文在解密后适用本授权书)学位论文作者签名:米露品 签字日期:2。*4月牛日导师签字:就卷:昉 签字日期:9年4月4日(C)1994-2023 China Academic Journal Electronic Publishing House.All rights reserved,http:/本文是兵团中青年领军人才项目“枣抗裂品种选育及抗裂
4、基因的挖掘”项目的 部分研究成果(项目编号:2018CB032)课题主持人:林敏娟教授(塔里木大学)本文是兵团南疆重点产业支撑计划项目“新疆梨、核桃、枣主要性状遗传资源评价(估)和功能 基因挖掘”项目的部分研究成果(项目编号:2017DB006)课题主持人:吴翠云教授(塔里木大学)(C)1994-2023 China Academic Journal Electronic Publishing House.All rights reserved,http:/CaCh处理对伏脆蜜枣裂果生理特性及 基因表达与代谢物的影响摘要伏脆蜜是早熟丰产,耐旱、耐储存、抗病、含糖量高,皮薄肉厚的鲜食品种,酸甜适
5、口,味道 极佳,深受消费者喜爱。但由于裂果率高,严重影响了果实经济价值。本文以易裂品种伏脆蜜 为试验材料,喷施外源CaCb,测定枣裂果关键时期(盛花后69d-90d)裂果率、果皮解剖结构、果 皮细胞壁代谢酶活性,对自然、清水、CaCb处理下枣盛花后69d、81d、90d枣果皮样品进行转录组 及代谢组学分析,筛选出与裂果相关基因及差异代谢物,为科学防治裂果提供理论依据。LCaCb处理枣果实裂果率在枣裂果期极显著低于自然及清水处理;喷施CaCb显著降低枣果皮 纤维素酶、果胶酶、SOD、POD酶活性,在盛花后84d-90d经CaCL处理的枣果皮CAT酶活性显著 高于自然及清水处理。CaCb处理后枣果
6、皮角质层较为完整,枣果皮角质层厚度、表皮层厚度、内表 皮层厚度、内表皮层细胞数显著增加,细胞排列紧密。2.利用Illumina平台对CaCl2处理样本进行转录组测序,与生物信息学方法相结合分析CaCl2处 理下的转录本水平变化。随着裂果时间的增加,CaCb处理子下差异表达基因数量呈先上升后下降的 变化趋势,裂果中期差异表达基因数量最多,通过GO及KEGG分析发现,CaCL处理下显著注释 的通路主要是糖类、苯丙烷类及脂质类等。其中CaCL处理下尸。42、加4基因表达量盛花后81d 低表达;CaCL处理下果胶酸裂解酶PMEIs41.CML、基因表达量在枣盛花期81d都 高于自然及清水处理;CaCb
7、处理下-Gq/3、GDSL-APG.XET1、XET2、等基因在裂果初期高表达,而GDSL-1等在CaCb处理下低表达;自然及清水处理下POD”基因表达量在高发期相比CaCb处 理快速上升;CaCb处理下果胶酯酶尸E3在裂果高发期下降幅度较大,说明这14个基因可能参与裂 果发生过程,14个基因在CaCl2处理下对枣裂果调控起到关键作用。3.利用非靶向代谢组学技术,研究CaCb处理下枣裂果不同时期枣果皮样品代谢物变化。结果 表明,在自然VSCaCb、清水VSCaCL两个比较组中发现32个差异代谢物,主要为糖类、氨基酸类、类黄酮及酯类。其中D-核糖、D-阿拉伯糖醇、海藻糖、半乳酸、五羟黄酮、表儿茶
8、素、L-天门冬酰(C)1994-2023 China Academic Journal Electronic Publishing House.All rights reserved,http:/胺、2-羟基肉桂酸、4-羟基肉桂酸、间香豆酸在CaCb处理下大量积累。KEGG结果表明,差异代谢 物注释在类黄酮生物合成、氨酰tRNA生物合成、磷酸戊糖途径、苯丙烷生物合成戊糖 和葡萄糖醛酸的相互转化等代谢通路。4.转录组及代谢组联合分析发现脂类代谢物及相关基因、苯丙烷代谢物及相关基因,碳水化合 物代谢物及相关基因,差异基因与差异代谢物相互作用共同调控伏脆蜜枣裂果,其中尸及 POD42基因与4-羟基肉
9、桂酸及2-羟基肉桂酸极显著相关。关键词:枣;裂果;生理指标;转录组;代谢组(C)1994-2023 China Academic Journal Electronic Publishing House.All rights reserved,http:/Effects of CaCh treatment on physiological characteristics,geneexpression and metabolites of fucuimi Jujube fruit crackingAbstractFucuimi is a fresh food variety with early
10、maturity and high yield,drought resistance,storage resistance,disease resistance,high sugar content,thin skin and thick meat.It is sour,sweet and palatable,with excellent taste,and is deeply loved by consumers.However,the high rate of fruit cracking has seriously affected the fruit q uality and yiel
11、d.In this paper,the cracking rate,peel anatomical structure and peel cell wall metabolic enzyme activity of jujube in the key period of fruit cracking(69d-90d after full bloom)were measured by spraying exogenous CaCh.The transcriptome and metabolomics of jujube peel samples treated with natural,clea
12、n water and CaCh at 69d,81d and 90d after full bloom were analyzed to screen the genes and differential metabolites related to fruit cracking,so as to provide a theoretical basis fbr scientific prevention and control of fruit cracking.l.The fruit cracking rate of CaCh Treatment was significantly low
13、er than that of natural and water treatment;Spraying CaCl?significantly reduced the activities of cellulase,pectinase,SOD and pod in jujube peel.The cat enzyme activity of jujube peel treated with CaCh was significantly higher than that of natural and clean water treatment from 84d to 90d after flow
14、ering.After CaCh treatment,the cuticle of jujube peel was relatively complete,the thickness of cuticle,epidennis,inner surface cortex and the number of cells in inner epidermis increased significantly,and the cells arranged closely.2.The transcriptome of CaCb treated samples was seq uenced by Illumi
15、na platform,and the changes of transcript level under CaCb treatment were analyzed by combining bioinfbrmatics methods.With the increase of fruit cracking time,the number of differentially expressed genes under CaCh treatment increased first and then decreased.The number of differentially expressed
16、genes was the largest in the middle of fruit cracking.Through GO and KEGG analysis,it was found that the pathways significantly annotated under CaCh treatment were mainly sugars,Phenylpropanes and lipids.The expression of POD42 and IAA14 genes under CaCh Treatment was low 81 days after full flowerin
17、g;The gene expressions of pectinate lyase PL5-2,PMEIs41,CML and GASAL in CaCh treatment were higher than those in natural and clean water treatment at 81 days of full flowering stage;Under CaCh treatment J3-GAL3,GDSL-APG,XET1,XET2 and other genes were highly expressed in the early stage of fruit cra
18、cking,while GDSL-1 and other genes were low expressed under CaCb treatment;Compared with CaCb treatment,the expression of POD 17 gene increased rapidly in high incidence period under natural and clear water treatment;Pectinesterase PE3 decreased significantly in the high incidence stage of fruit cra
19、cking under CaCh treatment,indicating that these 14 genes may be involved in the process of fruit cracking,and 14 genes play a key role in the regulation of fruit cracking under Ca treatment.3.Using non targeted metabonomics technology,the changes of metabolites in jujube peel sam pies at different
20、stages of jujube cracking under CaCh treatment were studied.The results showed that 3 2 different metabolites were found in the two comparison groups of natural vs CaCb and cle ar water vs CaCh,mainly sugars,amino acids,flavonoids and esters.D-Ribose,D-Arabitol,Trehal ose,Galactaric acid,Tricetin,Ep
21、icatechin,L-asparagine,2-hydroxycinnamic acid,4-hydroxycinnamic acid and m-coumaric acid were specifically expressed under CaCb treatment.KEGG results show ed that the differential metabolites were annotated in Flavonoid biosynthesis,Aminoacyl-tRNA bio(C)1994-2023 China Academic Journal Electronic P
22、ublishing House.All rights reserved,http:/synthesis,Pentose phosphate pathway,Phenylpropanoid biosynthesis,Pentose and glucuronate inte rconversions and other metabolic pathways.4.the combined analysis of transcriptome and metabolome showed that lipid metabolites and r elated genes,phenylpropane met
23、abolites and related genes,carbohydrate metabolites and related gen es,and the interaction between differential genes and differential metabolites jointly regulated the f ruit cracking of fuzui honey jujube.Among them,POD 17 and POD42 genes were significantly cor related with 4-hydroxycinnamic acid
24、and 2-hydroxycinnamic acid.Key words:Jujube;Dehiscent fruit;Physiological indicators;Transcriptome;Metabolomic(C)1994-2023 China Academic Journal Electronic Publishing House.All rights reserved,http:/目录第1章绪论.11.1 裂果研究现状.11.2 果实裂果影响因素.21.2.1 品种对裂果影响.21.2.2 枣果实的结构与裂果的关系.21.2.3 矿质元素对裂果的影响.31.2.4 细胞代谢酶对
25、裂果的影响.31.2.5 果皮细胞壁结构物质对裂果的影响.41.2.6 组学在植物与裂果研究中的应用.41.3 研究意义及内容.6第2章 试验材料与方法.72.1 试验材料与仪器设备.72.1.1 试验材料.72.1.2 试验试剂及仪器设备.72.2 试验方法.72.2.1 试验设计.72.2.2 田间裂果率调查.82.2.3 细胞壁代谢酶活性的测定.82.2.4 细胞壁物质含量测定.92.2.5 石蜡切片的制作.102.2.6 转录组测序.102.2.7 代谢组测序.142.3 数据处理.14第3章结果与分析.153.1 钙对伏脆蜜裂果生理特性的影响.153.1.1 CaCb对伏脆蜜裂果率的
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