Genetic Analysis and Preliminary Mapping of Seed Hull-less Gene Cphl in Zucchini

doi:10.7668/hbnxb.20195485

Abstract

Abstract:

The naked seeds of zucchinii possess significant natural advantages in processing.To investigate the genetic mechanisms underlying the hull-less trait in zucchini,we utilized the hulled 17pu10 (P₁) and the hull-less 17pu08 (P₂) as parental strains,and constructed populations of F₁ (P₁×P₂),F₂,and BC₁.The phenotypes of the zucchini seeds in the progeny population were assessed.The results showed that the number ratio of hulled seeds and hull-less seeds in the progeny population was in line with 3∶1 separation ratio.This indicated that the hull-less trait was regulated by a single gene and that the hull-less allele was recessive.Genetic mapping within this interval revealed that the gene for hull-less in zucchini was located between the markers InDel3157329 and InDel3724121,with genetic distances of 1.4,2.6 cM,respectively,while the physical distance was 0.6 Mb.The annotation and function analysis of 24 genes in the interval showed that 4 genes were directly or indirectly involved in the biosynthesis of cell wall,cellulose and lignin.Further analysis of the expression differences of 4 genes showed that only Cp4.1LG12g04350 and Cp4.1LG12g04370 had significant differences in expression levels during seed development.It was inferred that Cp4.1LG12g04350 or Cp4.1LG12g04370 was the candidate gene controlling the hull-less trait.In addition,InDel markers linked to hull-less genes were developed,which could be used as markers to identify zucchini hull-less traits,so as to accelerate the breeding of high-quality zucchini seed hull-less varieties.

Key words: Zucchini, Hull-less gene, Genetic analysis, Gene mapping, Gene linkage, InDel marked

DONG Chenchen, LIU Zehui, CAO Aiwan, XU Xiaoyong, LEI Fengjin, LIU Qinghua. Genetic Analysis and Preliminary Mapping of Seed Hull-less Gene Cphl in Zucchini[J]. Acta Agriculturae Boreali-Sinica, 2025, 40(2): 40-48. doi: 10.7668/hbnxb.20195485.

/ / Recommend / Download Citations

URL: http://www.hbnxb.net/EN/10.7668/hbnxb.20195485

http://www.hbnxb.net/EN/Y2025/V40/I2/40

Figures/Tables 10

References 35

[1]	申琼, 武峻新. 南瓜、西葫芦薄种皮形成机制研究进展[J]. 山西农业科学, 2014, 42(11): 1233-1235,1238. doi:10.3969/j.issn.1002-2481.2014.11.23.
	Shen Q, Wu J X. Study progress on the formation mechanism of hull-less seed in Cucurbita moschata and Cucurbita pepo[J]. Journal of Shanxi Agricultural Sciences, 2014, 42(11):1233-1235,1238.
[2]	张仲保. 无种壳西葫芦某些性状遗传规律的研究[J]. 西北农业学报, 2004, 13(3):93-96.doi:10.3969/j.issn.1004-1389.2004.03.024.
	Zhang Z B. Studies on the heredity of some characters in seed shell-less of squash[J]. Acta Agriculturae Boreali-occidentalis Sinica, 2004, 13(3):93-96.
[3]	Xue L, Yu G S, Gui M Z, Cui C S, Qu S P. Histological analysis of hulled and hull-less squash seed coat[J]. International Journal of Agriculture and Biology, 2013, 15(5):1047-1450.doi:10.1590/brag.2013.029.
[4]	Shen Q, Wu L Y, Zhao X H, Li Y L. Comparative analysis of electron microscopic structure of seed testa,lignin and biosynthesis-related enzyme activities in hulled and hull-less seeds of Cucurbita moschata[J]. Scientia Horticulturae, 2019, 245:137-143.doi:10.1016/j.scienta.2018.10.005.
[5]	申琼. 薄种皮南瓜种皮发育生理及分子标记筛选[D]. 太谷: 山西农业大学, 2019. doi:10.27285/d.cnki.gsxnu.2019.000078.
	Shen Q. Development physiology and molecular marker screening of Cucurbita moschata seed coat[D]. Taigu: Shanxi Agricultural University, 2019.
[6]	屈淑平, 秦俊芬, 靳楠楠, 桂明珠, 崔崇士. 美洲南瓜种皮发育的比较解剖学观察[J]. 园艺学报, 2011, 38(8):1547-1552.
	Qu S P, Qin J F, Jin N N, Gui M Z, Cui C S. Comparison of testa development in normal and hull-less strains of Cucurbita pepo[J]. Acta Horticulturae Sinica, 2011, 38(8):1547-1552.
[7]	张国华, 田慧颖, 何国庆. 益生菌安全性与功能性评价方法研究进展[J]. 山西农业科学, 2022, 50(11):1505-1510.doi:10.3969/j.issn.1002-2481.2022.11.03.
	Zhang G H, Tian H Y, He G Q. Research progress on evaluation methods of probiotic safety and function[J]. Journal of Shanxi Agricultural Sciences, 2022, 50(11):1505-1510.
[8]	Nakic' S N, Rade D, Škevin D, Štrucelj D, Mokrovcˇak Ž, Bartolic' M. Chemical characteristics of oils from naked and husk seeds of Cucurbita pepo L.[J]. European Journal of Lipid Science and Technology, 2006, 108(11):936-943.doi:10.1002/ejlt.200600161.
[9]	刘战霞, 吴宏, 郭慧静, 李斌斌, 吴洪斌, 贾文婷, 杨慧. 不同烘制预处理方式下裸仁南瓜籽烘烤工艺的比较[J]. 现代食品科技, 2023, 39(1):213-221.doi:10.13982/j.mfst.1673-9078.2023.1.0280.
	Liu Z X, Wu H, Guo H J, Li B B, Wu H B, Jia W T, Yang H. Comparison of roasting process of naked pumpkin seeds under different roasting pretreatment methods[J]. Modern Food Science and Technology, 2023, 39(1):213-221.
[10]	赵夏云, 杨静, 马关鹏, 瞿飞, 田颖, 文林宏. 裸仁南瓜种皮发育研究进展[J]. 中国瓜菜, 2021, 34(1):9-14.doi:10.16861/j.cnki.zggc.2021.0002.
	Zhao X Y, Yang J, Ma G P, Qu F, Tian Y, Wen L H. Research progress on seed coat development of hull-less seed in Cucurbita[J]. China Cucurbits and Vegetables, 2021, 34(1):9-14.
[11]	周祥麟. 裸仁南瓜的育成及其遗传规律的探讨[J]. 山西农业科学, 1983, 11(12):11-12.
	Zhou X L. Study on the breeding and genetic law of naked pumpkin[J]. Journl of Shanxi Agricultural Sciences, 1983, 11(12):11-12.
[12]	周祥麟. 裸仁南瓜的育成及其遗传性研究[J]. 园艺学报, 1987, 14(2):115-118.
	Zhou X L. A study on the breeding of naked kernel pumpkin and its genetic behavior[J]. Acta Horticulturae Sinica, 1987, 14(2):115-118.
[13]	刘恩璞, 魏淑琴, 王静华. 80831裸仁西葫芦的选育[J]. 中国蔬菜, 1989(2):6-8.doi:10.19928/j.cnki.1000-6346.1989.02.004.
	Liu E P, Wei S Q, Wang J H. Breeding of 80831 naked pumpkin[J]. China Vegetables, 1989(2):6-8.
[14]	徐东辉, 崔崇士, 张耀伟. 南瓜优势育种及遗传规律的研究进展[J]. 东北农业大学学报, 2004, 35(5):612-615.doi:10.3969/j.issn.1005-9369.2004.05.022.
	Xu D H, Cui C S, Zhang Y W. Advances in heterosis breeding and heredity law of pumpkin[J]. Journal of Northeast Agricultural University, 2004, 35(5):612-615.
[15]	薛应钰, 师桂英, 徐秉良, 陈荣贤. 裸仁美洲南瓜种皮性状遗传规律研究[J]. 草业学报, 2011, 20(3):205-210.
	Xue Y Y, Shi G Y, Xu B L, Chen R X. A study on the genetic law of seed coat characters in hull-less Cucurbita pepo[J]. Acta Prataculturae Sinica, 2011, 20(3):205-210.
[16]	李智媛, 屈淑平, 崔崇士. 美洲南瓜裸仁基因的AFLP和SCAR分子标记分析[J]. 东北农业大学学报, 2011, 42(7):67-71.doi:10.19720/j.cnki.issn.1005-9369.2011.07.012.
	Li Z Y, Qu S P, Cui C S. Development of AFLP and SCAR markers linked to hull-less n gene in pumpkin(Cucurbita pepo L.)[J]. Journal of Northeast Agricultural University, 2011, 42(7):67-71.
[17]	Gong L, Stift G, Kofler R, Pachner M, Lelley T. Microsatellites for the genus Cucurbita and an SSR-based genetic linkage map of Cucurbita pepo L.[J]. Theoretical and Applied Genetics, 2008, 117(1):37-48.doi:10.1007/s00122-008-0750-2. pmid: 18379753
[18]	武峻新. 南瓜遗传多样性及裸仁、矮生基因分子标记[D]. 太原: 山西大学, 2011. doi:10.7666/d.Y2682821.
	Wu J X. Assessment of genetic diversity and characterization of naked gene and dwarf gene in Cucurbita moschata using molecular makers[D]. Taiyuan: Shanxi University, 2011.
[19]	Zraidi A, Stift G, Pachner M, Shojaeiyan A, Gong L, Lelley T. A consensus map for Cucurbita pepo[J]. Molecular Breeding, 2007, 20(4):375-388.doi:10.1007/s11032-007-9098-6.
[20]	胡玉璐, 姜百灵, 陈凌, 王海岗, 曹晓宁, 王瑞云, 乔治军. 利用SSR创建中国北方黍稷资源DNA身份证[J]. 山西农业科学, 2022, 50(4):478-485.doi:10.3969/j.issn.1002-2481.2022.04.05.
	Hu Y L, Jiang B L, Chen L, Wang H G, Cao X N, Wang R Y, Qiao Z J. Using SSR to create DNA identity cards of broomcorn millet resources in northern China[J]. Journal of Shanxi Agricultural Sciences, 2022, 50(4):478-485.
[21]	Montero-Pau J, Blanca J, Bombarely A, Ziarsolo P, Esteras C, Martí-Gómez C, Ferriol M, Gómez P, Jamilena M, Mueller L, Picó B, Ca$\dot{n}$izares J. De ovo assembly of the zucchini genome reveals a whole-genome duplication associated with the origin of the Cucurbita genus[J]. Plant iotechnology ournal, 2018, 16(6):1161-1171.doi:10.1111/pbi.12860.
[22]	Takagi H, Abe A, Yoshida K, Kosugi S, Natsume S, Mitsuoka C, Uemura A, Utsushi H, Tamiru M, Takuno S, Innan H, Cano L M, Kamoun S, Terauchi R. QTL-seq:rapid mapping of quantitative trait loci in rice by whole genome resequencing of DNA from two bulked populations[J]. The Plant Journal, 2013, 74(1):174-183.doi:10.1111/tpj.12105.
[23]	Wyatt L E, Strickler S R, Mueller L A, Mazourek M. An acorn squash (Cucurbita pepo ssp.ovifera) fruit and seed transcriptome as a resource for the study of fruit traits in Cucurbita[J]. Horticulture Research, 2015, 2:14070.doi:10.1038/hortres.2014.70.
[24]	薛应钰, 师桂英, 徐秉良, 陈荣贤. 美洲南瓜(Cucurbita pepo)种皮发育形态观察及其相关酶活性测定[J]. 草业学报, 2011, 20(2):23-30.
	Xue Y Y, Shi G Y, Xu B L, Chen R X. Studies on morphology of seed coat development and its related enzyme activity assay in Cucurbita pepo[J]. Acta Prataculturae Sinica, 2011, 20(2):23-30.
[25]	孟秋峰, 汪炳良, 王毓洪, 孙崇德, 李永华. 园艺植物孢粉和种皮分类研究进展[J]. 长江蔬菜, 2008(20):1-3.doi:10.3865/j.issn.1001-3547.2008.20.001.
	Meng Q F, Wang B L, Wang Y H, Sun C D, Li Y H. Progress of classification for palynology and seed-coat of horticultural plant[J]. Journal of Changjiang Vegetables, 2008(20):1-3.
[26]	Meru G, Fu Y Q, Leyva D, Sarnoski P, Yagiz Y. Phenotypic relationships among oil,protein,fatty acid composition and seed size traits in Cucurbita pepo[J]. Scientia Horticulturae, 2018, 233:47-53.doi:10.1016/j.scienta.2018.01.030.
[27]	Murovec J, Drašlar K, Bohanec B. Detailed analysis of Cucurbita pepo seed coat types and structures with scanning electron microscopy[J]. Botany, 2012, 90(11):1161-1169.doi:10.1139/b2012-088.
[28]	邹伶威. 美洲南瓜裸仁基因的SSR分子标记[D]. 哈尔滨: 黑龙江大学, 2013. doi:10.7666/d.Y2323914.
	Zou L W. SSR molecular markers of naked kernel gene of Curcurbita pepo L.[D]. Harbin: Helongjiang University, 2013.
[29]	Meru G, Fu Y Q, Shrestha S, Michael V N, Dorval M, Mainviel R. Genomic position and markers associated with the hull-less seed trait in pumpkin[J]. Plants, 2022, 11(9):1238.doi:10.3390/plants11091238.
[30]	Kaur B, Garcha K S, Bhatia D, Khosa J S, Sharma M, Mittal A, Verma N, Dhatt A S. Identification of single major QTL and candidate gene(s) governing hull-less seed trait in pumpkin[J]. Frontiers in Plant Science, 2022, 13:948106.doi:10.3389/fpls.2022.948106.
[31]	Chen J H, Tao F, Xue Y Y, Xu B L, Li X W. Genome-wide identification of the WRKY gene family and functional characterization of CpWRKY5 in Cucurbita pepo[J]. International Journal of Molecular Sciences, 2024, 25(8):4177.doi:10.3390/ijms25084177.
[32]	Lyu X L, Shi L, Zhao M, Li Z P, Liao N Q, Meng Y Q, Ma Y Y, Zhou Y L, Xue Q, Hu Z Y, Yang J H, Zhang M F. A natural mutation of the NST1 gene arrests secondary cell wall biosynthesis in the seed coat of a hull-less pumpkin accession[J]. Horticulture Research, 2022,9:uhac136.doi:10.1093/hr/uhac136.
[33]	Shen Q, Weng Y Q. Alternative splicing of NAC transcription factor gene CmNST1 is associated with naked seed mutation in pumpkin,Cucurbita moschata[J]. Genes, 2023, 14(5):962.doi:10.3390/genes14050962.
[34]	Bezold T N, Mathews D, Loy J B, Minocha S C. Molecular analysis of the hull-less seed trait in pumpkin:expression profiles of genes related to seed coat development[J]. Seed Science Research, 2005, 15(3):205-217.doi:10.1079/ssr2005211.
[35]	Zhong R Q, Demura T, Ye Z H. SND1,a NAC domain transcription factor,is a key regulator of secondary wall synthesis in fibers of Arabidopsis[J]. The Plant Cell, 2006, 18(11):3158-3170.doi:10.1105/tpc.106.047399.

引物名称 Primer name	正向引物序列(5'—3') Forward primer(5'—3')	反向引物序列(5'—3') Reverse primer(5'—3')
Indel 1434239	AAAGGCGTAGTTTTCATAGT	TTGTGCAAAATCACCTTCAA
Indel 1447763	AGATGAGCTAAGGCCCAACA	TTACTGTGCCGAATGGTTTG
Indel 1476103	CACTTTCTGCTTACACCCAT	CCTCCGCTGTTTCTTCCTTC
Indel 2024961	AGTTGAAGAAGTGCTTGCTG	ATTGGTAGGAGACCTTTTGG
Indel 2124842	AATTAAATCAGCCACCACTA	CACAGTTCTACAACGGCAAG
Indel 2737845	ATGCACATCACCCCAAACAA	TATTAGTAACGGGTTAGATT
Indel 2864940	CATGTCACAGGGGATGTAGG	CTCGCAATTTATTTCTAAGGAT
Indel 2884389	CACGGCTGAAGATAGCATTT	AAGGGTCTCATACATTACAT
Indel 3157329	ACAAGAACCCCACCACATCA	TGCGAAGAAAACAAACAGAACA
Indel 3248462	AAGTCCCACATCGGCTAATT	CCTTTTCCAACTTTCGCTTA
Indel 3257054	GATAAGCATTCGACGTAGGC	GGTCATCCTCTAGGCAGCAT
Indel 3278372	ATTAGCGGAAAGATTATGAG	GAGTCACCTTTGATGTCTCA
Indel 3279258	GGAGGATAAATACCCAAACT	TACATTACCACCATTCCAAA
Indel 3478990	TGGTCGCTTGTATCCTGTTT	CTTATGTATGCCTATGTGAT
Indel 3680002	AAAACACGGTTAAACCACAT	GACGACAAGCGTACACTCAC
Indel 3724121	AGGTCCTTCCTATCACGATT	GAGGGTCTCACATTGCATAA
Indel 5674271	ACCCGACGTTTTAGTCCTTA	CACCCCGAACCATATCTTAT

引物名称 Primer name	正向引物序列(5'—3') Forward primer(5'—3')	反向引物序列(5'—3') Reverse primer(5'—3')
Indel 1434239	AAAGGCGTAGTTTTCATAGT	TTGTGCAAAATCACCTTCAA
Indel 1447763	AGATGAGCTAAGGCCCAACA	TTACTGTGCCGAATGGTTTG
Indel 1476103	CACTTTCTGCTTACACCCAT	CCTCCGCTGTTTCTTCCTTC
Indel 2024961	AGTTGAAGAAGTGCTTGCTG	ATTGGTAGGAGACCTTTTGG
Indel 2124842	AATTAAATCAGCCACCACTA	CACAGTTCTACAACGGCAAG
Indel 2737845	ATGCACATCACCCCAAACAA	TATTAGTAACGGGTTAGATT
Indel 2864940	CATGTCACAGGGGATGTAGG	CTCGCAATTTATTTCTAAGGAT
Indel 2884389	CACGGCTGAAGATAGCATTT	AAGGGTCTCATACATTACAT
Indel 3157329	ACAAGAACCCCACCACATCA	TGCGAAGAAAACAAACAGAACA
Indel 3248462	AAGTCCCACATCGGCTAATT	CCTTTTCCAACTTTCGCTTA
Indel 3257054	GATAAGCATTCGACGTAGGC	GGTCATCCTCTAGGCAGCAT
Indel 3278372	ATTAGCGGAAAGATTATGAG	GAGTCACCTTTGATGTCTCA
Indel 3279258	GGAGGATAAATACCCAAACT	TACATTACCACCATTCCAAA
Indel 3478990	TGGTCGCTTGTATCCTGTTT	CTTATGTATGCCTATGTGAT
Indel 3680002	AAAACACGGTTAAACCACAT	GACGACAAGCGTACACTCAC
Indel 3724121	AGGTCCTTCCTATCACGATT	GAGGGTCTCACATTGCATAA
Indel 5674271	ACCCGACGTTTTAGTCCTTA	CACCCCGAACCATATCTTAT

世代或组合 Generation & Combination	有壳株数 Number of hull seed	无壳株数 Number of hull-less seed	比例 Separate ratio	χ²值 χ² value
P₁(17pu10)	30	0
P₂(17pu08)	0	30
F₁(P₁×P₂)	48	0
F₂(F₁×F₁)	37	15	2.5∶1.0	0.23
BC₁(F₁×P₂)	159	181	1.00∶1.14	0.72

世代或组合 Generation & Combination	有壳株数 Number of hull seed	无壳株数 Number of hull-less seed	比例 Separate ratio	χ²值 χ² value
P₁(17pu10)	30	0
P₂(17pu08)	0	30
F₁(P₁×P₂)	48	0
F₂(F₁×F₁)	37	15	2.5∶1.0	0.23
BC₁(F₁×P₂)	159	181	1.00∶1.14	0.72

样品编号 Sample ID	原始测序数据量/Gb Amount of raw sequencing data	有效数据的数据量/Gb Amount of valid sequencing data	有效reads 所占比例/% Account for valid reads	质量值大于等于20的碱基所占比例(Q20)/% Account for quality values greater than or equal to 20 bases	质量值大于等于 30的碱基所占比例(Q30)/% Account for quality values greater than or equal to 30 bases	GC含量/% GC content
Hull mix	10.67	9.05	84.76	94.27	89.18	38.86
Hull-less mix	10.95	8.61	78.61	93.37	87.85	38.81
P₁	7.60	5.94	78.24	92.86	87.46	40.72
P₂	7.03	5.95	84.66	94.11	89.14	40.56

Please choose a citation manager

Content to export