葡萄Fe-S簇装配基因的鉴定、克隆和表达特征分析

doi:10.3864/j.issn.0578-1752.2021.23.012

中国农业科学 ›› 2021, Vol. 54 ›› Issue (23): 5068-5082.doi: 10.3864/j.issn.0578-1752.2021.23.012

葡萄Fe-S簇装配基因的鉴定、克隆和表达特征分析

张璐^1,²(),宗亚奇^1,²,徐维华²,韩蕾¹,孙浈育^1,²,陈朝晖³,陈松利³,张凯¹,程杰山¹,唐美玲^1,^2,³(),张洪霞¹,宋志忠¹()

¹鲁东大学农林工程研究院/山东省高等学校重点实验室-作物高产抗逆分子模块育种实验室,山东烟台 264025
²烟台农业科学研究院葡萄研究所,山东烟台 264000
³招远市大户庄园农林专业合作社,山东烟台 264000

收稿日期:2021-02-08 接受日期:2021-05-05 出版日期:2021-12-01 发布日期:2021-12-06
通讯作者: 唐美玲,宋志忠
作者简介:张璐,Tel：0535-6352051;E-mail： 1169464953@qq.com。
基金资助:
国家现代农业产业技术体系建设专项(CARS-29-16);山东省农业良种工程基金(2019LZGC009-2);烟台市科技计划(2018XSCC043);烟台市科技计划(2018XSCC043);国家重点研发计划(2019YFD1000500);烟台市葡萄与葡萄酒局专项资金(50012305073)

Identification, Cloning, and Expression Characteristics Analysis of Fe-S Cluster Assembly Genes in Grape

ZHANG Lu^1,²(),ZONG YaQi^1,²,XU WeiHua²,HAN Lei¹,SUN ZhenYu^1,²,CHEN ZhaoHui³,CHEN SongLi³,ZHANG Kai¹,CHENG JieShan¹,TANG MeiLing^1,^2,³(),ZHANG HongXia¹,SONG ZhiZhong¹()

¹The Engineering Research Institute of Agriculture and Forestry, Ludong University/Key Laboratory of Molecular Module-Based Breeding of High Yield and Abiotic Resistant Plants in Universities of Shandong, Yantai 264025, Shandong
²Institute of Grape, Yantai Academy of Agricultural Science, Yantai 264000, Shandong
³Zhaoyuan Dahu Manor Agriculture and Forestry Professional Cooperative, Yantai 264000, Shandong

Received:2021-02-08 Accepted:2021-05-05 Online:2021-12-01 Published:2021-12-06
Contact: MeiLing TANG,ZhiZhong SONG

摘要/Abstract

摘要：

【目的】从葡萄中克隆并鉴定Fe-S簇装配基因,在转录水平探索其组织特异性表达特征及其对缺铁胁迫的差异响应,明确主效基因。【方法】通过同源克隆法,在葡萄基因组中筛选并鉴定参与Fe-S簇装配的基因;借助生物信息学软件分析葡萄Fe-S簇装配相关基因及其编码蛋白的详细特征;利用实时荧光定量PCR分析Fe-S簇装配相关基因在葡萄不同组织部位的表达模式及其对缺铁胁迫的响应情况;利用MEGE 7.0软件建立不同植物ISU1同源蛋白的系统进化树。【结果】在葡萄基因组中检索并克隆获得46个Fe-S簇装配基因,分布于16条染色体上,含有1—21个长度不一的内含子,且主要分布于质体、线粒体和细胞质,分别含有14、21和11个基因成员;葡萄Fe-S簇装配蛋白在多种亚细胞结构中均有定位,且不同装配机制中蛋白的亚细胞定位情况差异很大;所选10种植物ISU1蛋白序列的一致性高达77%,系统发育树分析表明同一属的ISU1同源蛋白如十字花科的拟南芥和盐芥、禾本科的水稻和短柄草、蔷薇科的桃和苹果,倾向于紧密聚在一起,但葡萄ISU1和番茄ISU1紧密聚集在一起;葡萄Fe-S簇装配基因在3年生‘马瑟兰’成年树体和组培幼苗不同组织中的表达水平差异较大,其中,ISU1整体水平的表达量最为丰富（尤其是成熟期果实中的表达量最高）,其次是HSCA1、ISA2、NFU2、SUFA和SUFB等基因,而SUFE2、NFS1、HSCA2、HSCA6、TAH18和CIA2在本研究所有葡萄组织中均未检测到表达量;在‘马瑟兰’幼苗中,葡萄Fe-S簇装配基因对缺铁处理较为敏感,所有基因至少在1个检测的组织部位对缺铁处理有响应,其中,22个基因的表达水平在所有检测组织中均受缺铁处理调控：根部Fe-S簇装配基因的表达水平易受缺铁胁迫诱导而上调,但地上部（茎和叶）Fe-S簇装配基因的表达水平易受缺铁胁迫抑制而下调。【结论】从葡萄中克隆并鉴定了46个Fe-S簇装配基因,分别定位于质体、线粒体和细胞质;葡萄Fe-S簇装配基因在三年生成年树体和组培幼苗不同组织中的表达水平差异较大,且在葡萄幼苗不同组织中的转录水平对缺铁胁迫的响应具有显著差异;ISU1在葡萄所有组织中的整体表达量较高;葡萄ISU1和番茄ISU1同源蛋白遗传进化距离最接近。

关键词: 葡萄, 铁代谢, 缺铁, Fe-S簇, 装配机制, 基因克隆及表达

Abstract:

【Objective】The aim of this study was to isolate and characterize grape Fe-S cluster assembly genes. The tissue-specific expression characteristics and differential response to iron depletion at the transcriptional level were analyzed to screen the dominant and candidate Fe-S cluster assembly genes in grape. 【Method】The Fe-S cluster assembly genes were screened and identified in grape genome by homologous cloning. The detailed characteristics of Fe-S cluster assembly genes and their encoded proteins in grape were analyzed by using a variety of bioinformatics software. The expression patterns of Fe-S cluster assembly genes in different tissues of grape and their corresponding responses to iron deficiency stress were analyzed by using real-time fluorescent quantitative PCR. The phylogenetic trees of ISU1 homologous proteins from different plants were established by using MEGE 7.0 software. 【Result】 In total, 46 Fe-S cluster assembly genes were retrieved and cloned from grape, which were located on a total of 16 chromosomes, containing 1-21 introns with different lengths. Grape Fe-S cluster assembly genes were mainly distributed in plastid, mitochondrion and cytosol, containing 14, 21 and 11 genes, respectively. Grape Fe-S cluster assembly proteins were found with a variety of subcellular structures. Moreover, the subcellular localization of Fe-S cluster assembly proteins with different assembly mechanisms was quite distinct. The sequence identity of ISU1 homologous proteins from 10 plant species was as high as 77%. Phylogenetic tree analysis indicated that ISU1 members belonging to the same genus, such as Arabidopsis and the llungiella of Cruciferae, rice and Brachypodium of Gramineae, and peach and apple of Rosaceae, were tended to be closely clustered together, while grape ISU1 was closely clustered with tomato ISU1. The expression levels of Fe-S cluster assembly genes were different among different tissues of 3-year-old adult tree and tissue culture seedling of Marselan grape, and the differences were significant. In particular, the expression level of ISU1 gene was the most abundant (especially in the mature fruit), followed by HSAC1, ISA2, NFU2, SUFA and SUFB, while the expression levels of SUFE2, NFS1, HSCA2, HSCA6, TAH18 and CIA2 were not detected in tested tissues of grape. Grape Fe-S cluster assembly genes were more sensitive to iron deficiency treatment, and all genes responded to iron deficiency treatment in at least one detected tissue sample. Notably, the expression level of 22 Fe-S cluster assembly genes were significantly regulated by iron deficiency in all tested tissues, and the expression levels in roots were likely to be up-regulated by iron deficiency, while the expression levels in shoots (stems and leaves) were prone to be down-regulated. 【Conclusion】46 Fe-S cluster assembly genes were cloned and identified in grape, which were located in plastid, mitochondrion and cytosol, respectively. The expression levels of Fe-S cluster assembly genes were significantly distinct among different tissues of 3-year-old adult tree and tissue culture seedlings, while the transcription levels in different seedlings tissues were significantly different in response to iron deficiency. The overall expression level of ISU1 gene was the highest in all tested tissues of grape. The genetic evolution distance between grape and tomato ISU1 homologous proteins was the closest.

Key words: grape, iron metabolism, iron depletion, Fe-S cluster, assembly mechanism, gene cloning and expression

张璐,宗亚奇,徐维华,韩蕾,孙浈育,陈朝晖,陈松利,张凯,程杰山,唐美玲,张洪霞,宋志忠. 葡萄Fe-S簇装配基因的鉴定、克隆和表达特征分析[J]. 中国农业科学, 2021, 54(23): 5068-5082.

ZHANG Lu,ZONG YaQi,XU WeiHua,HAN Lei,SUN ZhenYu,CHEN ZhaoHui,CHEN SongLi,ZHANG Kai,CHENG JieShan,TANG MeiLing,ZHANG HongXia,SONG ZhiZhong. Identification, Cloning, and Expression Characteristics Analysis of Fe-S Cluster Assembly Genes in Grape[J]. Scientia Agricultura Sinica, 2021, 54(23): 5068-5082.

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链接本文: https://www.chinaagrisci.com/CN/10.3864/j.issn.0578-1752.2021.23.012

https://www.chinaagrisci.com/CN/Y2021/V54/I23/5068

图/表 12

表1

葡萄Fe-S簇装配基因CDS扩增所需引物"

基因Gene	上游引物 Forward primer (5′-3′)	下游引物 Reverse primer (5′-3′)	产物长度 Product length (bp)
ADX1	ATGTTCGCTTCTAGACTTTCA	TTAGTGTGGTTTTGGAATAAA	597
ADXR	ATGGCGTTTTGTCGTGCGAAG	TCATTCCATGGTAACTTTCAG	1461
ATM3	ATGGCGGGCGTTGCTTCTTGC	TCACGCCTCCAATTTAATGGT	2181
CIA1	ATGGATTTCTCGGAAGAGGGC	TCAGGTTATAGAAGCCAGCTC	1035
CIA2	ATGTTTTTCCCTGAGGTGGAC	TTAGAGTTCATTGGAGTAGAG	360
CIA3	ATGTTCTTAAATCATATTAGA	TCAACCATATGATGGAGCCAG	363
DRE2	ATGTTGCAAAATAGCACATTG	TTAAATGTCAGCCACAAGAA	819
ERV1	ATGTCTGAAACCCCATTTCAT	TTAATGTGATCTCTCACCAAA	597
FH	ATGGCTTCTGCTTCTTCTTCC	TTAGGAAAGGCTGATGGGTGT	594
GRXS14	ATGCTTATGCTTATATCAATA	TCAGGAGCACATCGCCTTCTC	552
GRXS15	ATGGCAAGGTCACTATCTAAC	TCATTCGGACTTTTCCTGGGG	513
GRXS16	ATGGCAACAATCAGTCTATCT	CTACTTCTTAAATAAACCAAC	894
HCF101	ATGAGGCTAACAAACACTCTT	TCATGCCTGTGCAGGAGTTAG	1671
HSCA1	ATGGCTGCCTCCGTTTTGCTC	CTACTTCTTCACCTCTTCATA	1842
HSCA2	ATGGCCACTTCGGTGCTTCTT	CTACTTCTTGGCCTGCTCATA	1716
HSCA3	ATGTCGGAGTGCCCAGCGATC	TCACATCACCATCCGGTCCCTT	1503
HSCA4	ATGTACGGTGCGTGGGGTCTA	CTACAGCTCATCATGGGAGTC	1947
HSCA5	ATGGACGAGGTGGGTGAGGA	TCACTTGCTCTCGGTGAAGTC	1677
HSCA6	ATGGAAAGCTCGTGGAGAAGA	TCAAAGTTCATCATGGGAATC	1821
HSCA7	ATGGCGGTGAAGAACAGGGCG	TCAGAACTGGAAGAAGCCCCA	1731
HSCB	ATGTGGAAGAAGAATCTATGG	TCAGAGTTTCTTCATGATTTC	804
IBA57	ATGGCCTCGCTTCTCAAGTCC	CTAAGCGGACGCAGTATCTTG	1113
INDL	ATGGAGTCACCGACCCAGGGA	TCACAGTGAAATCTCTGGCCG	1026
ISA1	ATGGCATCTTCATTTCTCGCG	CTACTTAGCAGCTCCAGGATT	402
ISA2	ATGCCCCAGCCCTCCACTTCG	TCACATTTCAGCAGCAAAGGA	453
ISA3	ATGTCAAGATCGGTGCTTCGG	CTATTCTTTCACCATGAAAGA	465
ISD11	ATGGCATCAACACCTTCGAAA	TTATTCCAACTGCCTTCTGGC	219
ISU1	ATGAAGCTTCAAATCAAGGTC	CTACTCAGTCTTTGCTCGCTT	273
MMS19	ATGGCACAACTGAGTCAATTG	TCAGAAATGAAGACTTCTGGA	3441
NAR1	ATGTCGGAGAAGTTCTCGGCA	TCACCAGTTGTGTAATTGAGA	1437
NBP35-1	ATGGAGAACGGCGACAGCAAT	TCAAGTGGTAGACAGTGCCTG	957
NBP35-2	ATGGAGTCACCGACCCAGGGA	TCACAGTGAAATCTCTGGCCG	1026
NFS1	ATGGCTTCCAGGCTTCATTCT	TTAGGTTATTTGTTTTTGTAG	3267
NFS2	ATGGCAAGTGTTCTGAAATT	CTACTTGAAAGAAGAGAAGA	1392
NFU1	ATGGCGTCTCTCACAGCAACA	CTAGTCACTGAAGACAACATT	642
NFU2	ATGCAAGGCGTGGTGGTGGCG	CTATAGAAGCTGAACTGCTGC	684
NFU3	ATGGGCTTCACAGCATTCACA	TTATCCATCTATCAACTGGAC	714
NFU4	ATGTTCATCCAAACTCAATCC	TTAATCCATCTGGCCAGATAG	609
SUFA	ATGCCCCAGCCCTCCACTTCG	TCACATTTCAGCAGCAAAGGA	453
SUFB	ATGGCTTCTCTACTGGCCAAC	CTAACCGACTGATCCTTCAAGC	1536
SUFC	ATGGCTCGATTCTGCTGCGCC	TTACAGTTGGGCCTCAGAAAT	966
SUFD	ATGGCTGCTACATTGCTTTCA	TCATGAGGAACCTTTAAGTGT	1443
SUFE1	ATGTCTCTGTCCAATCTCCAA	TCAACATAGATTCCCAAGTTG	1224
SUFE2	ATGGACTCTGCAACTTTGGG	TCATATCCTGAAAGGACTCTG	855
SUFE3	ATGGGGTGTACGGCGCAGGTG	TCAACTTATTGAAGTTGATC	1806
TAH18	ATGAATGGGAGGGAGAAGCAG	TCAAGACCATGCTTCCACATG	1899

表1

表2

本研究所用特异性表达引物"

基因 Gene	上游引物 Forward primer (5′-3′)	下游引物 Reverse primer (5′-3′)	产物长度 Product length (bp)
ADX1	AACCACAATCGGGGACCAAG	GTGGAACAGGCAAGTGAACC	250
ADXR	TGGGCATCCAGACTGCAAAA	TGGTCCACGTCTTCCTACCA	211
ATM3	CACCGACACCCTAGTTGGAC	TAGCCCCAACCAAGAAACCC	232
CIA1	GCAACGTTCTCCCTCCACTT	GACCCGGATGCATTCCAAGA	231
CIA2	TGTTTTTCCCTGAGGTGGACAT	GCCAATTACTGTTGCCATGCT	164
CIA3	AAACTTCTGCGCAGTTTGCC	GGAGCCAGACATTCATCAACC	167
DRE2	ATAGCACATTGGCCCTCACA	TGTCCCACCAGGCTTCAAAA	248
ERV1	TCGGCAGCAGAAGAAGGATG	CCCCACCTTGCATCAACTCT	222
FH	GGCCGCCATTGATTACAGTTC	GCTCACAGGAGAGGACAACC	235
GRXS14	GGAACCAAGGAGTTCCCACA	CGCCTTCTCCAACTGTTCCT	243
GRXS15	TGAAAGGGGTGCCTGATGTG	TTGTGGAAATGTGGGCCAGT	158
GRXS16	AAACTGAGCCGATTCCCCTG	CTCTGGCACCGATTTCCTGT	167
HCF101	GAGTGGGGAGAGCTGGACTA	AACAACAGCAACACAGGGGA	192
HSCA1	CTCGCTTCCCCTTCTCTCTC	CCACTGATGGTGTAGTCCGAG	235
HSCA2	CCATTCAGCTCAAAGCCTGC	GGGAGCTCTGAGTTTAGTTCCA	243
HSCA3	AGAGAGGTTTGCAAAGGAGGA	GTCCCTTTCCTGATGACTTTCC	244
HSCA4	ACCAATGACAAGGAGGAAGACA	CACTCGAGGGCCTCTTTCAC	176
HSCA5	CAGAGACTGCCGAGAAAGCA	GCGTTGAACCACCAACAAGG	246
HSCA6	GTCAAGGCTGAGGACAAGGG	GACAGCTTCCACCTCATCAATC	165
HSCA7	TACACGAGTTTGCAGAAGAGG	AGCCCCACCACTCCTCATAAA	177
HSCB	TCCATGTTCCTCAACTGCCC	ACAAGAGTCGCAAGCCAGAA	181
IBA57	GTGGATGCCACTGTCTTGGA	ATGTGCCAGCAGGATCAACA	202
INDL	AGTGGTTCTGCGGGGTTTAG	ATCAGCATCAAGCACACCCA	168
ISA1	CCTAACCCTAACGGACGCTG	CCAATGACATGCATGAGAGCC	213
ISA2	GCCCTCCACTTCGGTTTCTC	CCAGAGCATCCACCCTGTTT	219
ISA3	TTGAGTGTCGAGACTGGTGG	TGCTCACCAGGAAAGCAGAA	193
ISD11	ACGCGATTTCTCGGACTACAAT	TATTCCAACTGCCTTCTGGC	156
ISU1	CCATCGCCTCTTCCTCTGTT	AGTCTTTGCTCGCTTGGCT	188
MMS19	AGCTTGCTGTTGGGAGTTGT	GGGTTTGAGGACGAACTGCT	231
NAR1	AAAGCTGAGGTTGTGACGGG	AAAGGAAGCCCTTGACTGGG	204
NBP35-1	TTCCTAGTGGTTGATGCCCC	ACACCTCACTGAAATCTGTCAC	232
NBP35-2	AGTGGTTCTGCGGGGTTTAG	ATCAGCATCAAGCACACCCA	168
NFS1	GACAAATCAAAAAGCCATTTTCCAA	CACGTACTGCTCTCGGGTT	150
NFS2	GTTCAGGTGGGGTTCCGTAG	AGGCTTCTGTGAGGTTGCTG	183
NFU1	CAAAACTCAGCGTTTCGCCT	ATTTGTGGGCCGAGTAGAGC	163
NFU2	CCTACTGCTGCAGAGCCAAA	GCAACAGCCTTGACAACTCG	172
NFU3	AAACACGCCCTAGAGCCTTC	AGCCTTGACAACTCGGCTAC	150
NFU4	GGGAAGCCACTGTTCCTTGA	CGATACTCGATGTCCCCACC	152
SUFA	GCCCTCCACTTCGGTTTCTC	CCAGAGCATCCACCCTGTTT	219
SUFB	ACAAAGTTTGCAGATGACAGGA	CTAGCCCCACTGCATAGACC	221
SUFC	CCATGCAGTCATGGGGAAGA	ACCTGGGATCTCTACTGGGG	190
SUFD	AATTGCCCACTGGCGTTTTC	ATATCCCCACCCTCAACGGA	217
SUFE1	TGGACATGCAGGTGTTAGGG	TTCCCAAGTTGATTTCCCTGTCT	219
SUFE2	TGATTCGGGTTTTGGACGGA	AGATATTGCAGTGCCAGAGACA	227
SUFE3	TCCTGAGGTCCAAGGTGTCT	GCATCTGCCAAAGAGCAACC	243
TAH18	GCGCTATTTCTTTGAGGCAAG	AATGGCATTTGCACTGAAGGG	174

表2

图1

表3

葡萄、拟南芥和桃Fe-S簇装配基因对比"

位置 Location	基因 Gene	登录号 Locus ID
位置 Location	基因 Gene	拟南芥 Arabidopsis	葡萄Grape	桃 Peach	水稻 Rice
质体 Plastid	NFS2	At1g08490	GSVIVT01037384001	ppa005298m	Os12g18900
	SUFE1	At4g26500	GSVIVT01014835001	ppa007330m	Os09g09790
	SUFE2	At1g67810	GSVIVT01000550001	ppa017530m	-
	SUFE3	At5g50210	GSVIVT01007621001	ppa001921m	Os12g19304
	SUFA	At1g10500	GSVIVT01022247001	ppa012351m	Os06g05400
	SUFB	At4g04770	GSVIVT01012742001	ppa003788m	Os01g61400
	SUFC	At3g10670	GSVIVT01036588001	-	Os03g21490
	SUFD	At1g32500	GSVIVT01013180001	ppa004982m	Os01g03650
	NFU1	At4g01940	GSVIVT01028158001	ppa011214m	Os03g20010
	NFU2	At5g49940	GSVIVT01023274001	ppa011050m	Os11g07916
	NFU3	At4g25910	GSVIVT01029242001	ppa010743m	Os06g47940
	HCF101	At3g24430	GSVIVT01015424001	ppa006650m	Os01g52170
	GRXS14	At3g54900	GSVIVT01032936001	ppa012220m	Os03g63420
	GRXS16	At2g38270	GSVIVT01011178001	ppa009373m	Os12g07650
线粒体 Mitochondria	NFS1	At5g65720	GSVIVT01003603001	ppa005512m	Os09g16910
	ISD11	At5g61220	GSVIVT01007919001	ppa013993m	Os08g14070
	ISU1	At4g22220	GSVIVT01013764001	ppa012356m	Os01g47340
	ISU2	At3g01020	-	-	Os05g49300
	ISU3	At4g04080	-	-	-
	ISA1	At2g16710	GSVIVT01033834001	ppa013203m	Os12g30030
	ISA2	At2g36260	GSVIVT01022247001	ppa12351m	Os01g01610
	ISA3	At5g03905	GSVIVT01033842001	ppa012679m	Os08g28230
	NFU4	At3g20970	GSVIVT01011272001	ppa009781m	Os05g06330
	NFU5	At1g51390	-	-	-
	ADX1	At4g21090	GSVIVT01015024001	ppa012568m	Os09g26650
	ADX2	At4g05450	-	ppa012238m	Os07g01930
	ADXR	At4g32360	GSVIVT01015352001	ppa004960m	Os02g17700
	FH	At4g03240	GSVIVT01000441001	ppa011940m	Os01g57460
	HSCA1	At4g37910	GSVIVT01038517001	ppa002402m	Os02g53420
	HSCA2	At5g09590	GSVIVT01006769001	ppa001973m	Os03g02260
	HSCA3	-	GSVIVT01008331001	ppa002222m	Os09g31486
	HSCA4	-	GSVIVT01038580001	ppa002489m	-
	HSCA5	-	GSVIVT01026014001	ppa002572m	-
	HSCA6	-	GSVIVT01019607001	-	-
	HSCA7	-	GSVIVT01031125001	-	-
	HSCB	At5g06410	GSVIVT01022555001	ppa016242m	Os12g27070
	INDL	At4g19540	GSVIVT01000762001	ppa019981m	Os03g42880
	IBA57	At4g12130	GSVIVT01004911001	ppa006632m	Os06g04380
	GRXS15	At3g15660	GSVIVT01017244001	ppa012405m	Os01g07950
细胞质 Cytosol	ATM3	At5g58270	GSVIVT01024527001	ppa002114m	Os06g03770
	ERV1	At1g49880	GSVIVT01037859001	ppa012227m	Os03g10850
	NAR1	At4g16440	GSVIVT01007214001	ppa005089m	Os03g53750
	NBP35-1	At5g50960	GSVIVT01001845001	ppa005998m	Os04g40880
	NBP35-2	-	GSVIVT01000762001	ppa007759m	-
	TAH18	At3g02280	GSVIVT01031054001	ppa002941m	Os01g53250
	DRE2	At5g18400	GSVIVT01010181001	ppa009994m	Os04g57810
	CIA1	At2g26060	GSVIVT01033839001	ppa007909m	Os07g14830
	CIA2	At1g68310	GSVIVT01018962001	ppa012624m	Os04g50864
	CIA3	-	GSVIVT01034035001	ppa012667m	-
	MMS19	At5g48120	GSVIVT01016980001	ppa023072m	Os07g08050

表3

表4

葡萄Fe-S簇装配基因信息"

位置 Location	基因 Gene	GenBank登录号 GenBank No.	染色体定位 Chromosome location	编码区 CDS (bp)	氨基酸数目 Amino acid No.	内含子数目 Intron No.
质体 Plastid	NFS2	GSVIVT01037384001	chr6:14614433..14627074 forward	1392	463	9
	SUFE1	GSVIVT01014835001	chr19:10332463..10337676 reverse	1224	407	5
	SUFE2	GSVIVT01000550001	chr1:7199706..7203084 forward	855	284	2
	SUFE3	GSVIVT01007621001	chr17:11153636..11159809 reverse	1806	601	6
	SUFA	GSVIVT01022247001	chr7:17815443..17819261 forward	453	150	2
	SUFB	GSVIVT01012742001	chr10:1165899..1176476 reverse	1536	511	3
	SUFC	GSVIVT01036588001	chr13:18723793..18744366 forward	966	321	5
	SUFD	GSVIVT01013180001	chr2:7019468..7043516 forward	1443	480	2
	NFU1	GSVIVT01028158001	chr7:4298897..4301308 forward	642	213	2
	NFU2	GSVIVT01023274001	chr12:20249550..20254467 reverse	684	227	3
	NFU3	GSVIVT01029242001	chr11:18133595..18136869 forward	714	237	3
	HCF101	GSVIVT01015424001	chr11:3539907..3550291 forward	1671	556	14
	GRXS14	GSVIVT01032936001	chr14:24341805..24359981 reverse	552	183	3
	GRXS16	GSVIVT01011178001	chr8:7822419..7828262 reverse	894	297	1
线粒体 Mitochondria	NFS1	GSVIVT01003603001	chrUn:11176164..11189530 forward	3267	1088	8
	ISD11	GSVIVT01007919001	chr17:7527950..7530932 reverse	219	72	1
	ISU1	GSVIVT01013764001	chr1:7813881..7823597 forward	273	90	2
	ISA1	GSVIVT01033834001	chr8:17357057..17361779 forward	402	133	2
	ISA2	GSVIVT01022247001	chr7:17815443..17819261 forward	453	150	2
	ISA3	GSVIVT01033842001	chr8:17302334..17306498 forward	465	154	3
	NFU4	GSVIVT01011272001	chr13:10306616..10320274 reverse	609	202	4
	ADX1	GSVIVT01015024001	chr11:532570..538242 forward	597	198	6
	ADXR	GSVIVT01015352001	chr11:2891542..2897423 reverse	1461	486	12
	FH	GSVIVT01000441001	chr12:8053062..8056733 reverse	594	197	4
	HSCA1	GSVIVT01038517001	chr3:10899748..10904643 forward	1842	613	7
	HSCA2	GSVIVT01006769001	chrUn:28290681..28295479 forward	1716	571	8
	HSCA3	GSVIVT01008331001	chr17:3189753..3194345 forward	1503	500	9
	HSCA4	GSVIVT01038580001	chr16:21738678..21742840 reverse	1947	648	8
	HSCA5	GSVIVT01026014001	chr18:25847096..25853527 forward	1677	558	7
	HSCA6	GSVIVT01019607001	chr2:1900757..1904873 forward	1821	606	8
	HSCA7	GSVIVT01031125001	chr14:1920620..1924241 forward	1731	576	8
	HSCB	GSVIVT01022555001	chr8:4909017..4918207 forward	804	267	5
	INDL	GSVIVT01000762001	chr7:498142..504281 reverse	1026	341	8
	IBA57	GSVIVT01004911001	chr2:4748585..4752156 forward	1113	370	4
	GRXS15	GSVIVT01017244001	chr9:5862839..5867495 reverse	513	170	5
细胞质 Cytosol	ATM3	GSVIVT01024527001	chr6:9155464..9213374 reverse	2181	726	19
	ERV1	GSVIVT01037859001	chr3:7051150..7055710 forward	597	198	7
	NAR1	GSVIVT01007214001	chrUn:30906448..30911807 forward	1437	478	10
	NBP35-1	GSVIVT01001845001	chr14:26263495..26271378 reverse	957	318	3
	NBP35-2	GSVIVT01000762001	chr7:498142..504281 reverse	1026	341	8
	TAH18	GSVIVT01031054001	chr14:21477906..21501706 reverse	1899	632	11
	DRE2	GSVIVT01010181001	chr1:16897462..16906512 reverse	819	272	6
	CIA1	GSVIVT01033839001	chr8:17322989..17332968 forward	1035	344	9
	CIA2	GSVIVT01018962001	chr4:18301283..18304437 reverse	360	119	5
	CIA3	GSVIVT01034035001	chr8:15682406..15687298 forward	363	120	3
	MMS19	GSVIVT01016980001	chr9:3133979..3155012 forward	3441	1146	21

表4

图2

表5

Fe-S簇装配基因亚细胞定位预测"

蛋白位置 Protein location	蛋白 Protein	亚细胞定位 Subcellular localization (%)
蛋白位置 Protein location	蛋白 Protein	叶绿体Chloroplast	线粒体Mitochondria	细胞质Cytosol	细胞核Nucleus	内质网膜 Endoplastic reticulum	液泡膜 Vacular membrane	细胞质膜 Plasma membrane	高尔基体 Golgi
质体 Plastid	NFS2	71.43	28.57	-	-	-	-	-	-
	SUFE1	7.14	-	21.43	57.15	-	7.14	7.14	-
	SUFE2	92.86	7.14	-	-	-	-	-	-
	SUFE3	7.14	-	64.29	14.29	7.14	-	7.14	-
	SUFA	85.72	7.14	7.14	-	-	-	-	-
	NFU1	72	28	-	-	-	-	-	-
	NFU2	85.72	-	-	7.14	7.14	-	-	-
	NFU3	57.15	-	21.43	7.14	-	-	14.28	-
	SUFB	57.50	32.50	-	10.00	-	-	-	-
	SUFC	85.71	14.29	-	-	-	-	-	-
	SUFD	68.3	21.95	-	9.75	-	-	-	-
	HCF101	57.14	14.29	-	-	-	21.43	-	7.14
	GRXS14	92.86	-	-	-	-	7.14	-	-
	GRXS16	83.72	16.28	-	-	-	-	-	-
线粒体 Mitochondria	NFS1	35.71	35.71	-	28.58	-	-	-	-
	ISD11	-	77.42	-	16.13	-	-	6.45	-
	ISU1	85.72	-	14.28	-	-	-	-	-
	ISA1	92.85	7.15	-	-	-	-	-	-
	ISA2	78.57	7.15	14.28	-	-	-	-	-
	ISA3	21.63	45.94	27.03	5.40	-	-	-	-
	NFU4	50	14.28	-	14.28	-	7.14	-	-
	ADX1	14.28	78.58	-	7.14	-	-	-	-
	ADXR	50	35.72	7.14	7.14	-	-	-	-
	FH	50	35.72	-	14.28	-	-	-	-
	HSCA1	14.28	85.72	-	-	-	-	-	-
	HSCA2	14.28	85.72	-	-	-	-	-	-
	HSCA3	21.42	-	78.58	-	-	-	-	-
	HSCA4	-	-	-	-	100	-	-	-
	HSCA5	7.14	7.14	78.58	-	-	-	7.14	-
	HSCA6	-	-	-	-	100	-	-	-
	HSCA7	43.76	-	15.62	15.62	-	12.5	-	12.5
	HSCB	71.44	14.28	-	14.28	-	-	-	-
	INDL	14.28	-	78.58	7.14	-	-	-	-
	IBA57	64.30	14.28	7.14	7.14	-	-	7.14	-
	GRXS15	42.86	57.14	-	-	-	-	-	-
细胞质 Cytosol	ATM3	7.14	7.14	-	-	28.57	-	57.15	-
	ERV1	14.28	-	14.28	57.16	-	7.14	7.14	-
	NAR1	21.42	7.14	42.88	21.42	-	7.14	-	-
	NBP35-1	57.16	-	21.42	21.42	-	-	-	-
	NBP35-2	14.28	-	78.58	7.14	-	-	-	-
	TAH18	7.14	-	35.71	57.15	-	-	-	-
	DRE2	21.42	-	21.42	57.16	-	-	-	7.14
	CIA1	14.28	-	28.58	53	-	7.14	-	-
	CIA2	7.14	-	64.30	21.42	-	-	-	7.14
	CIA3	-	-	100	-	-	-	-	-
	MMS19	14.28	-	28.57	35.73	-	14.28	-	7.14

表5

图3

图4

表6

图5

图6

参考文献 24

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基因 Gene	拟南芥 Arabidopsis thaliana	盐芥 Thellungiella halophila	短柄草 Brachypodium sylvaticum	水稻 Rice	番茄 Tomato	柑橘 Orange	苹果 Apple	桃 Peach	葡萄 Grape
ISU1	At4g22220	Thhalv10026418m	Bra020855	Os01g47340	Solyc03g112900	orange1.1g030644m	MDP0000778166	ppa012356m	GSVIVT01013764001
ISU2	At3g01020	Thhalv10028164m	Bra013601	Os05g49300	Solyc07g007450	-	-	-	-
ISU3	At4g04080	Thhalv10029418m	Bra029483	-	-	-	-	-	-
NFU1	At4g01940	Thhalv10028839m	Bra000905	Os03g20010	Solyc01g079220	orange1.1g027469m	MDP0000245391	ppa011214m	GSVIVT01028158001
NFU2	At5g49940	Thhalv10014572m	Bra037947	Os11g07916	Solyc01g103710	orange1.1g026830m	MDP0000285539	ppa011050m	GSVIVT01023274001
NFU3	At4g25910	Thhalv10026125m	Bra013933	Os06g47940	Solyc05g044630	orange1.1g038446m	MDP0000952041	ppa010743m	GSVIVT01029242001
NFU4	At3g20970	Thhalv10021272m	Bra031245	Os05g06330	Solyc11g007120	orange1.1g023823m	MDP0000150995	ppa009781m	GSVIVT01011272001
NFU5	At1g51390	Thhalv10011711m	Bra018906	-	-	-	-	-	-

葡萄Fe-S簇装配基因的鉴定、克隆和表达特征分析

Identification, Cloning, and Expression Characteristics Analysis of Fe-S Cluster Assembly Genes in Grape

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