西瓜果形基因图位克隆及分子标记开发

doi:10.3864/j.issn.0578-1752.2022.14.011

中国农业科学 ›› 2022, Vol. 55 ›› Issue (14): 2812-2824.doi: 10.3864/j.issn.0578-1752.2022.14.011

西瓜果形基因图位克隆及分子标记开发

段雅如¹(),高美玲^1,²(),郭宇¹,梁晓雪¹,刘秀杰³,徐洪国¹,刘继秀³,高越³,栾非时⁴

1.齐齐哈尔大学生命科学与农林学院,黑龙江齐齐哈尔 161006
2.黑龙江省抗性基因工程与寒地生物多样性保护重点实验室,黑龙江齐齐哈尔 161006
3.齐齐哈尔市农业技术推广中心,黑龙江齐齐哈尔 161006
4.东北农业大学园艺园林学院,哈尔滨 150030

收稿日期:2021-11-03 接受日期:2022-02-03 出版日期:2022-07-16 发布日期:2022-07-26
联系方式: 段雅如,E-mail： 1141941808@qq.com。
基金资助:
国家自然科学基金(31972437);国家自然科学基金(31772334);国家自然科学基金(31401891);齐齐哈尔大学研究生创新科研项目(YJSCX2021025)

Map-Based Cloning and Molecular Marker Development of Watermelon Fruit Shape Gene

DUAN YaRu¹(),GAO MeiLing^1,²(),GUO Yu¹,LIANG XiaoXue¹,LIU XiuJie³,XU HongGuo¹,LIU JiXiu³,GAO Yue³,LUAN Feishi⁴

1. College of Life Sciences, Agriculture and Forestry, Qiqihar University, Qiqihar 161006, Heilongjiang
2. Heilongjiang Provincial Key Laboratory of Resistance Gene Engineering and Preservation of Biodiversity in Cold Areas, Qiqihar 161006, Heilongjiang
3. Qiqihar Agricultural Technology Extension Center, Qiqihar 161006, Heilongjiang
4. College of Horticulture and Landscape Architecture, Northeast Agricultural University, Harbin 150030

Received:2021-11-03 Accepted:2022-02-03 Published:2022-07-16 Online:2022-07-26

摘要/Abstract

摘要：

【目的】基于GBS（Genotyping-by-sequencing）高密度遗传图谱初步定位结果,通过图位克隆方法对西瓜果形基因进行精细定位,并开发功能性分子标记,有助于全面研究果形基因的功能及分子标记辅助选择育种。【方法】利用114份纯合西瓜品系全基因组重测序数据及其果形指数表型进行GWAS（genome-wide association study）分析,结合GBS高密度遗传图谱初步定位结果确定果形基因候选区段,以商业小型西瓜品种纯化所得品系K2（椭圆形、FSI=1.54±0.13）和L1（圆形、FSI=1.11±0.07）为材料构建F₂群体,通过开发分子标记对果形基因ClFSI进行精细定位,根据西瓜参考基因组‘97103’v1注释信息确定候选基因,并通过实时荧光定量PCR（qRT-PCR）进行验证。【结果】利用1 152份F₂群体,最终将ClFSI精细定位于3号染色体的FMFSI-1与FMFSI-2标记之间物理距离约63 kb区间内,共包含5个注释基因。其中Cla011257属于已报道与控制果实纵径和果形相关的SUN基因家族。经测序分析发现,西瓜椭圆形品系K2在该基因第3外显子上存在两个非同义突变位点Chr3：26846636 G-A和Chr3：26847041 G-A（‘97103’v1）,分别导致天冬酰胺（Asn）替换为天冬氨酸（Asp）和谷氨酸（Glu）替换为赖氨酸（Lys）,并利用Chr3：26847041突变位点开发功能性分子标记FSICAPS-2。qRT-PCR分析表明,K2（椭圆形）与Charleston Gray（细长形）中候选基因表达量无显著性差异,但均显著高于L1（圆形）。【结论】本研究将控制西瓜果形的ClFSI精细定位于3号染色体63 kb区间内,推测Cla011257为最终目的基因,Chr3：26847041和Chr3：26846636突变位点是导致果形不同程度伸长的重要位点,并开发了可以同时鉴定Cla011257多种突变类型的功能标记FSICAPS-2。

关键词: 西瓜, 果形基因, 图位克隆, 分子标记

Abstract:

【Objective】 Based on the result of fruit shape gene preliminary mapping by genotyping-by-sequencing (GBS) high-density genetic map, the fine mapping of the fruit shape gene was conducted by using map-based cloning method, and the functional molecular markers were developed in watermelon. The present study could facilitate comprehensive study on the function of fruit shape gene and molecular marker-assisted selection breeding. 【Method】GWAS (Genome-wide association study) analysis was performed on 114 watermelon inbred lines by using whole-genome re-sequencing data and their fruit shape index phenotypes. The candidate regions of fruit shape gene ClFSI were confirmed by combing the results of GWAS with fruit shape gene preliminary mapping. The F₂ segregating population were derived from a cross between two inbred lines K2 (oval, FSI=1.54±0.13) and L1 (round, FSI=1.11±0.07), which were purified from commercial small watermelon varieties. Fine-mapping of the fruit shape gene ClFSI was conducted by developing molecular markers. Candidate gene was identified by gene annotation of the candidate region in watermelon reference genome ‘97103’ v1 and validated by using real-time quantitative PCR (qRT-PCR). 【Result】 The 1 152 F₂ plants were used for fine mapping and the ClFSI gene was finally mapped into 63 kb region on chromosome 3 between FMFSI-1 and FMFSI-2, containing a total of 5 annotated genes. The Cla011257 gene belonged to the SUN gene family that has been reported to control the fruit shape. There were two SNPs identified in the genomic region of third exon of ClFSI gene in the watermelon oval line K2. One SNP was a mutation from G to A at Chr3: 26846636 (‘97103’ v1), which resulted in a mutation from asparagine acid (Asn) to aspartic acid (Asp). Another SNP was a mutation from G to A at Chr3: 26846636 (‘97103’ v1), which resulted in another mutation from glutamic acid (Glu) to lysine acid (Lys). The FSICAPS-2 functional molecular marker was developed based on the Chr3: 26847041 SNP site. qRT-PCR expression analysis showed that candidate gene expression level was not significantly difference in K2 (oval) and Charleston Gray (elongated), but both were significantly higher than that in L1 (round). 【Conclusion】In this study, the ClFSI gene was mapped into a 63 kb candidate region on chromosome 3, and Cla011257 was the good candidate gene. Chr3: 26847041 and Chr3: 26846636 mutations were important loci that caused elongation of different degrees in fruit shape. A functional marker FSICAPS-2 was developed, which could identify multiple mutation types of Cla011257 simultaneously.

Key words: watermelon, fruit shape gene, map-based cloning, molecular marker

段雅如,高美玲,郭宇,梁晓雪,刘秀杰,徐洪国,刘继秀,高越,栾非时. 西瓜果形基因图位克隆及分子标记开发[J]. 中国农业科学, 2022, 55(14): 2812-2824.

DUAN YaRu,GAO MeiLing,GUO Yu,LIANG XiaoXue,LIU XiuJie,XU HongGuo,LIU JiXiu,GAO Yue,LUAN Feishi. Map-Based Cloning and Molecular Marker Development of Watermelon Fruit Shape Gene[J]. Scientia Agricultura Sinica, 2022, 55(14): 2812-2824.

图/表 15

图1

表1

表2

表3

图2

表 4

图3

图4

表5

表6

表7

图5

图 6

图7

图8

参考文献 37

[1]	HARTMAN J L, PERKINS V P, WEHNER T C. Citrulline and arginine are moderately heritable in two red-fleshed watermelon populations. HortScience, 2019, 54(2): 200-205. doi: 10.21273/HORTSCI13715-18
[2]	RODRÍGUEZ G R, MUÑOS S, ANDERSON C, SIM S C, MICHEL A, CAUSSE M, GARDENER B B M, FRANCIS D, VAN DER KNAAP E. Distribution of SUN, OVATE, LC, and FAS in the tomato germplasm and the relationship to fruit shape diversity. Plant Physiology, 2011, 156(1): 275-285. doi: 10.1104/pp.110.167577. doi: 10.1104/pp.110.167577
[3]	WANG Y P, CLEVENGER J P, ILLA-BERENGUER E, MEULIA T, VAN DER KNAAP E, SUN L. A comparison of sun, ovate, fs8.1 and auxin application on tomato fruit shape and gene expression. Plant and Cell Physiology, 2019, 60(5): 1067-1081. doi: 10.1093/pcp/pcz024. doi: 10.1093/pcp/pcz024
[4]	LIU J P, VAN ECK J, CONG B, TANKSLEY S D. A new class of regulatory genes underlying the cause of pear-shaped tomato fruit. PNAS, 2002, 99(20): 13302-13306. doi: 10.1073/pnas.162485999. doi: 10.1073/pnas.162485999
[5]	XIAO H, JIANG N, SCHAFFNER E, STOCKINGER E J, VAN DER KNAAP E. A retrotransposon-mediated gene duplication underlies morphological variation of tomato fruit. Science, 2008, 319(5869): 1527-1530. doi: 10.1126/science.1153040. doi: 10.1126/science.1153040
[6]	ZHU W Y, HUANG L, CHEN L, YANG J T, WU J N, QU M L, YAO D Q, GUO C L, LIAN H L, HE H L, PAN J S, CAI R. A high-density genetic linkage map for cucumber (Cucumis sativus L.): Based on specific length amplified fragment (SLAF) sequencing and QTL analysis of fruit traits in cucumber. Frontiers in Plant Science, 2016, 7: 437. doi: 10.3389/fpls.2016.00437. doi: 10.3389/fpls.2016.00437
[7]	GAO Z H, ZHANG H Y, CAO C X, HAN J, LI H, REN Z H. QTL mapping for cucumber fruit size and shape with populations from long and round fruited inbred lines. Horicultural Plant Journal, 2020, 6(3): 132-144.
[8]	PAN Y P, LIANG X J, GAO M L, LIU H Q, MENG H W, WENG Y Q, CHENG Z H. Round fruit shape in WI7239 cucumber is controlled by two interacting quantitative trait loci with one putatively encoding a tomato SUN homolog. Theoretical and Applied Genetics, 2017, 130(3): 573-586. doi: 10.1007/s00122-016-2836-6. doi: 10.1007/s00122-016-2836-6
[9]	栾非时, 矫士琦, 盛云燕, 朱子成. 甜瓜果实相关性状QTL分析. 东北农业大学学报, 2017, 48(3): 1-9. doi: 10.19720/j.cnki.issn.1005-9369.2017.03.001. doi: 10.19720/j.cnki.issn.1005-9369.2017.03.001
	LUAN F S, JIAO S Q, SHENG Y Y, ZHU Z C. Mapping of QTL for fruit traits in melon. Journal of Northeast Agricultural University, 2017, 48(3): 1-9. doi: 10.19720/j.cnki.issn.1005-9369.2017.03.001. (in Chinese) doi: 10.19720/j.cnki.issn.1005-9369.2017.03.001
[10]	王岭, 才羿, 王桂超, 王迪, 盛云燕. 甜瓜SLAF图谱构建及果实相关性状QTL分析. 中国农业科学, 2021, 54(19): 4196-4206. doi: 10.3864/j.issn.0578-1752.2021.19.014. doi: 10.3864/j.issn.0578-1752.2021.19.014
	WANG L, CAI Y, WANG G C, WANG D, SHENG Y Y. Specific length amplified fragment (SFLA) sequencing mapping construction and QTL analysis of fruit related traits in muskmelon. Scientia Agricultura Sinica, 2021, 54(19): 4196-4206. doi: 10.3864/j.issn.0578-1752.2021.19.014. (in Chinese) doi: 10.3864/j.issn.0578-1752.2021.19.014
[11]	WEETMAN L M. Inheritance and correlation of shape, size and color in the watermelon. Iowa agr.expt.sta.res.bul, 1937, 228(2): 342-350.
[12]	POOLE C F, GRIMBALL P C. Interaction of sex, shape, and weight genes in watermelon. Journal of Agricultural Research, 1944, 71: 533-552.
[13]	TANAKA T, WIMOL S, MIZUTABI T. Inheritance of fruit shape and seed size of watermelon. Journal of the Japanese Society for Horticultural Science, 1995, 64(3): 543-548.
[14]	GUNER N, WEHNER T C. The genes of watermelon. Hortscience A Publication of the American Society for Horticultural Science, 2004, 39(6): 1175-1182.
[15]	LOU L L, WEHNER T C. Qualitative inheritance of external fruit traits in watermelon. Hortence A Publication of the American Society for Horticultural Science, 2016, 51(5): 487-496.
[16]	SANDLIN K, PROTHRO J, HEESACKER A, KHALILIAN N, OKASHAH R, XIANG W W, BACHLAVA E, CALDWELL D G, TAYLOR C A, SEYMOUR D K, WHITE V, CHAN E, TOLLA G, WHITE C, SAFRAN D, GRAHAM E, KNAPP S, MCGREGOR C. Comparative mapping in watermelon [Citrullus lanatus (thunb.) matsum. et nakai]. Theoretical and Applied Genetics, 2012, 125(8): 1603-1618. doi: 10.1007/s00122-012-1938-z. doi: 10.1007/s00122-012-1938-z
[17]	CHENG Y, LUAN F S, WANG X Z, GAO P, ZHU Z C, LIU S, BALOCH A M, ZHANG Y S. Construction of a genetic linkage map of watermelon (Citrullus lanatus) using CAPS and SSR markers and QTL analysis for fruit quality traits. Scientia Horticulturae, 2016, 202: 25-31. doi: 10.1016/j.scienta.2016.01.004
[18]	REN Y, MCGREGOR C, ZHANG Y, GONG G Y, ZHANG H Y, GUO S G, SUN H H, CAI W T, ZHANG J, XU Y. An integrated genetic map based on four mapping populations and quantitative trait loci associated with economically important traits in watermelon (Citrullus lanatus). BMC Plant Biology, 2014, 14: 33. doi: 10.1186/1471-2229-14-33. doi: 10.1186/1471-2229-14-33
[19]	REDDY U K, ABBURI L, ABBURI V L, SAMINATHAN T, CANTRELL R, VAJJA V G, REDDY R, TOMASON Y R, LEVI A, WEHNER T C, NIMMAKAYALA P. A genome-wide scan of selective sweeps and association mapping of fruit traits using microsatellite markers in watermelon. Journal of Heredity, 2014, 106(2): 166-176. doi: 10.1093/jhered/esu077. doi: 10.1093/jhered/esu077
[20]	KIM K H, HWANG J H, HAN D Y, PARY M, KIM S, CHOI D, KIM Y, LEE G P, KIM S T, PARK Y H. major quantitative trait loci and putative candidate genes for powdery mildew resistance and fruit-related traits revealed by an intraspecific genetic map for watermelon (Citrullus lanatus var. lanatus). PLoS ONE, 2015, 10(12): e0145665.
[21]	卢丙洋, 周慧文, 陈欣, 栾非时, 王学征, 姜羽. 西瓜果实几个性状的QTL分析. 果树学报, 2016, 33(10): 1206-1218. doi: 10.13925/j.cnki.gsxb.20160002. doi: 10.13925/j.cnki.gsxb.20160002
	LU B Y, ZHOU H W, CHEN X, LUAN F S, WANG X Z, JIANG Y. QTL analysis of fruit traits in watermelon. Journal of Fruit Science, 2016, 33(10): 1206-1218. doi: 10.13925/j.cnki.gsxb.20160002. (in Chinese) doi: 10.13925/j.cnki.gsxb.20160002
[22]	刘传奇, 高鹏, 栾非时. 西瓜遗传图谱构建及果实相关性状QTL分析. 中国农业科学, 2014, 47(14): 2814-2829. doi: 10.3864/j.issn.0578-1752.2014.14.012. doi: 10.3864/j.issn.0578-1752.2014.14.012
	LIU C Q, GAO P, LUAN F S. Construction of a genetic linkage map and QTL analysis of fruit-associated traits in watermelon. Scientia Agricultura Sinica, 2014, 47(14): 2814-2829. doi: 10.3864/j.issn.0578-1752.2014.14.012. (in Chinese) doi: 10.3864/j.issn.0578-1752.2014.14.012
[23]	DOU J L, ZHAO S J, LU X, HE N, ZHANG L, ALIi A, KUANG H H, LIU W G. Genetic mapping reveals a candidate gene (ClFS1) for fruit shape in watermelon (Citrullus lanatus L.). Theoretical and Applied Genetics, 2018, 131(4): 947-958. doi: 10.1007/s00122-018-3050-5
[24]	MARAGAL S, RAO E S, LAKSHMANA REDDY D C. Genetic analysis of fruit quality traits in prebred lines of watermelon derived from a wild accession of Citrullus amarus. Euphytica, 2019, 215(12): 1-15. doi: 10.1007/s10681-019-2527-x. doi: 10.1007/s10681-019-2527-x
[25]	LEGENDRE R, KUZY J, MCGREGOR C. Markers for selection of three alleles of ClSUN25-26-27a (Cla011257) associated with fruit shape in watermelon. Molecular Breeding, 2020, 40(2): 1-13. doi: 10.1007/s11032-020-1104-2. doi: 10.1007/s11032-020-1104-2
[26]	李娜, 尚建立, 李楠楠, 周丹, 孔胜楠, 王吉明, 马双武. 西瓜果实形状的分子精准鉴定. 园艺学报, 2021, 48(7): 1386-1396. doi: 10.16420/j.issn.0513-353x.2021-0152. doi: 10.16420/j.issn.0513-353x.2021-0152
	LI N, SHANG J L, LI N N, ZHOU D, KONG S N, WANG J M, MA S W. Accurate molecular identification for fruit shape in watermelon (Citrullus lanatus). Acta Horticulturae Sinica, 2021, 48(7): 1386-1396. doi: 10.16420/j.issn.0513-353x.2021-0152. (in Chinese) doi: 10.16420/j.issn.0513-353x.2021-0152
[27]	NEFF M M, TURK E, KALISHMAN M. Web-based primer design for single nucleotide polymorphism analysis. Trends in Genetics, 2002, 18: 613-615. doi: 10.1016/S0168-9525(02)02820-2
[28]	UNTERGASSER A, CUTCUTACHE I, KORESSAAR T, YE J, FAIRCLOTH B C, REMM M, ROZEN S G. Primer3: new capabilities and interfaces. Nucleic Acids Research, 2012, 40(15): e115. doi: 10.1093/nar/gks596. doi: 10.1093/nar/gks596
[29]	LI Y H, YANG L M, PATHAK M, LI D W, HE X M, WENG Y Q. Fine genetic mapping of cp: A recessive gene for compact (dwarf) plant architecture in cucumber, Cucumis sativus L. Theoretical and Applied Genetics, 2011, 123(6): 973-983. doi: 10.1007/s00122-011-1640-6. doi: 10.1007/s00122-011-1640-6
[30]	SANGER F, NICKLEN S, COULSON A R. DNA sequencing with chain-terminating inhibitors. Proceedings of the National Academy of Sciences of the United States of America, 1977, 74(12): 5463-5467.
[31]	KONG Q S, YUAN J X, GAO L Y, ZHAO L Q, CHENG F, HUANG Y, BIE Z L. Evaluation of appropriate reference genes for gene expression normalization during watermelon fruit development. PLoS ONE, 2015, 10(6): e0130865. doi: 10.1371/journal.pone.0130865. doi: 10.1371/journal.pone.0130865
[32]	PAN Y P, WANG Y H, MCGREGOR C, LIU S, LUAN F S, GAO M L, WENG Y Q. Genetic architecture of fruit size and shape variation in cucurbits: A comparative perspective. Theoretical and Applied Genetics, 2020, 133(1): 1-21. doi: 10.1007/s00122-019-03481-3. doi: 10.1007/s00122-019-03481-3
[33]	CLEVENGER J P, VAN HOUTEN J, BLACKWOOD M, RODRÍGUEZ G R, JIKUMARU Y, KAMIYA Y, KUSANO M, SAITO K, VISA S, VAN DER KNAAP E. Network analyses reveal shifts in transcript profiles and metabolites that accompany the expression of SUN and an elongated tomato fruit. Plant Physiology, 2015, 168(3): 1164-1178. doi: 10.1104/pp.15.00379. doi: 10.1104/pp.15.00379
[34]	MONFORTE A J, DIAZ A, CAÑO-DELGADO A, VAN DER KNAAP E. The genetic basis of fruit morphology in horticultural crops: Lessons from tomato and melon. Journal of Experimental Botany, 2014, 65(16): 4625-4637. doi: 10.1093/jxb/eru017. doi: 10.1093/jxb/eru017
[35]	WU S, XIAO H, CABRERA A, MEULIA T, VAN DER KNAAP E. SUN regulates vegetative and reproductive organ shape by changing cell division patterns. Plant Physiology, 2011, 157(3): 1175-1186. doi: 10.1104/pp.111.181065. doi: 10.1104/pp.111.181065
[36]	HUANG Z J, HOUTEN J, GONZALEZ G, XIAO H, KNAAP E. Genome-wide identification, phylogeny and expression analysis of SUN, OFP and YABBY gene family in tomato. Molecular Genetics and Genomics, 2013, 288(3/4): 111-129. doi: 10.1007/s00438-013-0733-0. doi: 10.1007/s00438-013-0733-0
[37]	JIANG N, GAO D Y, XIAO H, WAN DER KNAAP E. Genome organization of the tomato sun locus and characterization of the unusual retrotransposon Rider. The Plant Journal, 2010, 60(1): 181-193. doi: 10.1111/j.1365-313X.2009.03946.x

引物名称Primer name	正向序列 Forward primer	反向序列 Reverse primer	产物长度 Product length (bp)
FSIgene-1	GGACTCAATTTGTAGAGCATGG	AAGAACGCAGTGATGAGCAA	460
FSIgene-2	TCGAGTTTGCTCATCACTGC	CGGAATTAAGCGACGACCTA	463
FSIgene-3	TCCGGCAATCTGGGATTT	ATTAGGCGAGGAAAGCACTG	556
FSIgene-4	CAGTGCTTTCCTCGCCTAAT	GAAAAATGGGGAGAGCAACC	547

引物名称 Primer name	正向序列 Forward primer	反向序列 Reverse primer
qClFSI	AAGGGTTTCGTTGAATGAATTG	GGCTTCTTGAACTTGAGAACA
Actin	CCATGTATGTTGCCATCCAG	GGATAGCATGGGGTAGAGCA

性状 Trait	K2 (n=20)	L1 (n=20)	F₁(n=20)	2016F₂
性状 Trait	均值±标准差 Mean±SD	均值±标准差 Mean±SD	均值±标准差 Mean±SD	均值±标准差 Mean±SD	范围 Range	峰度 Kurtosis	偏度 Skewness
果实纵径 Fruit length (cm)	20.4±3.0A	15.3±2.4B	19.3±2.6A	16.2±2.6	10.8-23.6	-0.18	0.40
果实横径 Fruit width (cm)	13.3±1.5b	13.8±1.8b	15.4±1.3a	13.6±1.2	9.7-18.5	-0.42	0.16
果形指数 Fruit shape index (cm)	1.54±0.13A	1.11±0.07B	1.26±0.22B	1.20±0.20	0.92-1.77	0.27	1.09

性状 Trait	果实纵径 Fruit length	果实横径 Fruit diameter	果形指数 Fruit shape index
果实纵径 Fruit length	1
果实横径 Fruit width	0.43**	1
果形指数 Fruit shape index	-0.38**	0.67**	1

引物名称 Primer name	物理位置（V1） Physical position	变异位点 Variation locus
FMFSI-3	chr3-26695927	C/A	Fa: GAAGGTGACCAAGTTCATGCTGTAAGAACAGCCAAATGTTGAACAAATC
			Fb: GAAGGTCGGAGTCAACGGATTTTGTAAGAACAGCCAAATGTTGAACAAATA
			R: CAAGACAGTGATCTTTTCACAACACAATAATT
FMFSI-4	chr3-26775520	G/C	Fa: GAAGGTGACCAAGTTCATGCTGTAATAAATGTGGAGGAAATATGGCTAC
			Fb: GAAGGTCGGAGTCAACGGATTGTAATAAATGTGGAGGAAATATGGCTAG
			R: TAACCATTTACTTTTTTGGTTTTGGTTTTTGAAAA
FMFSI-1	chr3-26801978	C/T	Fa: GAAGGTGACCAAGTTCATGCTACACATTCTAATGCCCTAGTTAGTAC
			Fb: GAAGGTCGGAGTCAACGGATTCACACATTCTAATGCCCTAGTTAGTAT
			R: TTGGATCATCGAATTTTATGAACACTCCTAT

西瓜果形基因图位克隆及分子标记开发

Map-Based Cloning and Molecular Marker Development of Watermelon Fruit Shape Gene

RichHTML

PDF

赞

可视化

摘要/Abstract

引用本文

使用本文

图/表 15

参考文献 37

相关文章 15

Metrics

本文评价

推荐阅读 0

引物名称 Primer name	物理位置（V1） Physical position	变异位点 Variation locus	正向序列 Forward sequence	反向序列 Reverse sequence	限制性内切酶Restriction endonuclease
GFSI-1	chr3-26846636	C/T	AGACTACTTCTCGATTCCATGAATT	CGGAATTTCCATGACTCTGA	EcoR I (dCPAS)
FSICAPS-2	chr3-26847041	C/T	ACCGCCAGAGTTCACAAATC	ATTAGGCGAGGAAAGCACTG	Taq I
FM-FSI-2	chr3-26864789	C/G	AATTGATGGGGGTATCGTGA	TGGTTGGGGTTATTTGGGTA	Rsa I
CW3CAPS47	chr3-26882166	T/G	GAACCAATACCAACGGGAAT	TGCATGAGCCAAAATTCACT	Xho I
CHR3-FSI-3	chr3-26940134	T/C	CTTTGAAGTTCGGAGGTTGG	ATGGATCAAACAGACGCAAA	Taq I
CHR3-FSI-5	chr3-26945758	A/G	CCAAGCAGGCCAAATAACAT	GCCAAATTTTCGAAGCACAG	Hind II
CHR3-FSI-7	chr3-27106460	G/A	CAAAAGCCTCCAAGTTCCAG	AAGCTGGTGGTCTTGGTTTG	Taq I
CHR3-FSI-11	chr3-27121470	C/T	TTGATCGCTGGAGATGAATG	GCATCGGTTTTGGAAGGATA	Hind III

编号 No.	基因号 Gene ID	基因功能 Gene function	基因位置 Region in ‘97103’ v1
1	Cla011252	低温诱导蛋白 Cold induced protein like (AHRD V1 *---Q94JH8_ORYSJ)	Chr3: 26806979-26807611 (+strand) 633 bp
2	Cla011253	精氨酸tRNA合成酶 Arginine tRNA synthetase (AHRD V1 **** O23247_ARATH); 含有Interpro结构域IPR001278 Contains interpro domains IPR001278,精氨酸tRNA合成酶 Arginine tRNA synthetase,IC类 Class IC	Chr3: 26809083-26816437 (-strand) 7355 bp
3	Cla011254	未知蛋白质 Unknown protein (AHRD V1)	Chr3: 26819389-26819613 (+strand) 225 bp
4	Cla011255	激酶家族蛋白 Kinase family proteins (AHRD V1 ***-D7MCM2_ARALL);含有Interpro结构域IPR002290 Contains interpro domains IPR002290,丝氨酸/苏氨酸蛋白激酶 Serine/ Threonine protein kinase	Chr3: 268219 46-26833394 (+strand) 11449 bp
5	Cla011256	果胶酯酶 Pectin esterase (AHRD V1 ***-B9RKG6_RICCO);含有Interpro结构域IPR000070 Contains interpro domains IPR000070,果胶酯酶 Pectin esterase,具有催化作用 With catalysis	Chr3: 26838409-26839804 (+strand) 1396 bp
6	Cla011257	钙调素结合家族蛋白 Calmodulin binding family proteins (AHRD V1 *- Q9ZU28_ ARATH);含有Interpro域IPR000048 IQ Contains interpro domains IPR000048 IQ,钙调蛋白结合区 Calmodulin-binding domain	Chr3: 26846490-26847637 (-strand) 1148 bp

[1]	侯普兴, 王勇, 冯俊涛, 马志卿, 吴华. 75种植物乙醇提取物对两种土传病原菌的抑制活性[J]. 中国农业科学, 2026, 59(2): 322-335.
[2]	吕涛, 孙国清, 郭栋材, 陈全家, 蔡永生, 樊标星, 曲延英, 郑凯. 海岛棉纤维强度QTL紧密连锁InDel分子标记开发及效应评价[J]. 中国农业科学, 2025, 58(9): 1684-1701.
[3]	贾玉静, 李超男, 潘志雄, 杨德龙, 毛新国, 景蕊莲. 小麦TaTIFY11c-4A的克隆及遗传效应分析[J]. 中国农业科学, 2025, 58(17): 3357-3371.
[4]	曾跃辉, 邹文广, 赵福铭, 肖长春, 黄建鸿, 马彬林, 杨旺兴, 韦新宇, 许旭明. 一个新的水稻颖花开裂基因OsSG2的图位克隆与功能验证[J]. 中国农业科学, 2025, 58(11): 2062-2080.
[5]	黄立强, 江如, 朱波汁, 彭焕, 许翀, 宋家雄, 陈敏, 李永青, 黄文坤, 彭德良. 马铃薯主栽品种抗马铃薯金线虫鉴定及抗性分子标记检测[J]. 中国农业科学, 2024, 57(8): 1506-1516.
[6]	关志林, 靳丰蔚, 刘婷婷, 王毅, 谭莹莹, 杨春慧, 李蕊彤, 王博, 刘克德, 董云. 甘蓝型油菜光叶性状的遗传分析及基因定位[J]. 中国农业科学, 2024, 57(4): 650-662.
[7]	吴传磊, 胡晓渝, 王伟, 苗龙, 白鹏宇, 王郭伋, 李娜, 舒阔, 邱丽娟, 王晓波. 大豆油分相关功能基因分子标记开发与鉴定及优异等位变异聚合分析[J]. 中国农业科学, 2024, 57(22): 4402-4415.
[8]	陈文杰, 陈渊, 韦清源, 汤复跃, 郭小红, 陈淑芳, 覃夏燕, 韦荣昌, 梁江. 利用高世代转录组测序挖掘控制南方大豆皱叶症候选基因[J]. 中国农业科学, 2024, 57(15): 2914-2930.
[9]	高福, 王睿, 刘东军, 孙会言, 王子叶, 宋维富, 李天亚. 黑龙江省88份小麦品种（系）抗秆锈基因鉴定及抗性评价[J]. 中国农业科学, 2024, 57(13): 2568-2582.
[10]	刘华, 曾凡佩, 王倩, 陈国权, 苗利娟, 秦利, 韩锁义, 董文召, 杜培, 张新友. 花生栽培种和野生种Arachis sp.30119六倍体杂种的创制与鉴定[J]. 中国农业科学, 2024, 57(10): 1870-1881.
[11]	周警卫, 叶博伟, 张朋飞, 张宇庆, 郝敏, 尹毓若, 袁婵, 李志康, 李顺达, 夏先春, 何中虎, 张宏军, 兰彩霞. 国内外153份小麦种质条锈病抗性鉴定与评价[J]. 中国农业科学, 2024, 57(1): 18-33.
[12]	白斌, 张怀志, 杜久元, 张晓洋, 何瑞, 伍玲, 张哲, 张耀辉, 曹世勤, 刘志勇. 西北条锈菌源区冬小麦育种抗条锈病基因的利用现状与策略[J]. 中国农业科学, 2024, 57(1): 4-17.
[13]	赵梓钧, 吴如会, 王硕, 张君, 游静, 段倩楠, 唐俊, 张新芳, 韦秘, 刘金艳, 李云峰, 何光华, 张婷. 水稻PDL2的突变导致小穗外稃退化[J]. 中国农业科学, 2023, 56(7): 1248-1259.
[14]	朱洪慧, 李映姿, 高远卓, 林泓, 王成洋, 晏紫仪, 彭瀚平, 李田野, 熊茂, 李云峰. 水稻短宽粒基因SWG1的图位克隆[J]. 中国农业科学, 2023, 56(7): 1260-1274.
[15]	臧新山, 王康文, 张先亮, 王雪平, 王军, 梁雨, 裴小雨, 任翔, 吕宇龙, 高宇, 王星星, 彭云玲, 马雄风. 棉花功能基因图位克隆的研究进展[J]. 中国农业科学, 2023, 56(23): 4635-4647.