数量性状位点揭示锌指蛋白CONSTANS-LIKE 4与西瓜柱头颜色相关

doi:10.1016/j.jia.2024.03.070

Journal of Integrative Agriculture ›› 2024, Vol. 23 ›› Issue (7): 2292-2305.DOI: 10.1016/j.jia.2024.03.070

数量性状位点揭示锌指蛋白CONSTANS-LIKE 4与西瓜柱头颜色相关

收稿日期:2023-05-18 接受日期:2023-10-25 出版日期:2024-07-20 发布日期:2024-07-01

Quantitative trait loci identification reveals zinc finger protein CONSTANS-LIKE 4 as the key candidate gene of stigma color in watermelon (Citrullus lanatus)

Shuang Pei¹, Zexu Wu¹, Ziqiao Ji¹, Zheng Liu², Zicheng Zhu^1#, Feishi Luan^1#, Shi Liu^1#

¹Key Laboratory of Biology and Genetic Improvement of Horticultural Crops (Northeast Region), Ministry of Agriculture and Rural Affairs/College of Horticulture and Landscape Architecture, Northeast Agricultural University, Harbin 150030, China
²Institute of Rural Revitalization Science and Technology, Heilongjiang Academy of Agricultural Sciences, Harbin 150028, China

Received:2023-05-18 Accepted:2023-10-25 Online:2024-07-20 Published:2024-07-01
About author:#Correspondence Zicheng Zhu, Tel: +86-451-55190593, E-mail: zzc1983sc@163.com; Feishi Luan, Tel: +86-451-55190836, E-mail: luanfeishi@neau.edu.cn; Shi Liu, Tel: +86-451-55190204, E-mail: shiliu@neau.edu.cn
Supported by:
This work was supported by fundings from the Natural Science Funds for Outstanding Youth of Heilongjiang Province, China (YQ2022C011), the National Natural Science Foundation of China (32172577), the China Agriculture Research System of MOF and MARA, China (CARS-25), the Taishan Industrial Leading Talents Project, China (LJNY202112), and the Natural Science Foundation of Heilongjiang Province, China (LH2022C025).

摘要/Abstract

摘要：

柱头颜色是西瓜重要的农艺性状，在传粉过程中起着重要作用。然而，迄今为止有关这方面的研究报道较少。本研究通过对ZXG1553（P₁，橙色柱头）和W1-17（P₂，黄色柱头）F₂群体遗传分析表明，柱头颜色是一个数量性状，黄色对橙色表现为显性。BSA-seq (Bulk seq analysis sequencing) 测序分析发现在6号染色体3.75 Mb的区域存在与柱头颜色相关的染色体片段，并在CAPS标记Chr06_8338913和Chr06_9344593之间检测到一个主效QTL位点Clqsc6.1，候选区间约1.01 Mb，包含51个注释基因。通过RNA-seq、实时荧光定量PCR和基因结构比对等分析，推测Cla97C06G117020 （Zinc finger protein CONSTANS-LIKE 4）为西瓜柱头颜色性状的候选基因，Cla97C06G117020在橙色柱头中表达量低于黄色柱头。利用9份西瓜材料重测序数据分析后发现，Cla97C06G117020编码区存在1个非同义SNP突变。柱头颜色的形成往往与类胡萝卜素有关，我们发现类胡萝卜素代谢途径中ClCHYB （Beta-carotene hydroxylase）的表达趋势与Cla97C06G117020一致，在橙柱头中表达量较少，推测Cla97C06G117020和ClCHYB可能存在相互作用影响柱头颜色形成，该研究为西瓜柱头颜色基因精细定位和调控机制提供了理论依据。

Abstract:

Stigma color is a critical agronomic trait in watermelon that plays an important role in pollination. However, there are few reports on the regulation of stigma color in watermelon. In this study, a genetic analysis of the F₂ population derived from ZXG1553 (P₁, with orange stigma) and W1-17 (P₂, with yellow stigma) indicated that stigma color is a quantitative trait and the orange stigma is recessive compared with the yellow stigma. Bulk segregant analysis sequencing (BSA-seq) revealed a 3.75 Mb segment on chromosome 6 that is related to stigma color. Also, a major stable effective QTL Clqsc6.1 (QTL stigma color) was detected in two years between cleaved amplified polymorphic sequencing (CAPS) markers Chr06_8338913 and Chr06_9344593 spanning a ~1.01 Mb interval that harbors 51 annotated genes. Cla97C06G117020 (annotated as zinc finger protein CONSTANS-LIKE 4) was identified as the best candidate gene for the stigma color trait through RNA-seq, quantitative real-time PCR (qRT-PCR), and gene structure alignment analysis among the natural watermelon panel. The expression level of Cla97C06G117020 in the orange stigma accession was lower than in the yellow stigma accessions with a significant difference. A nonsynonymous SNP site of the Cla97C06G117020 coding region that causes amino acid variation was related to the stigma color variation among nine watermelon accessions according to their re-sequencing data. Stigma color formation is often related to carotenoids, and we also found that the expression trend of ClCHYB (annotated as β-carotene hydroxylase) in the carotenoid metabolic pathway was consistent with Cla97C06G117020, and it was expressed in low amounts in the orange stigma accession. These data indicated that Cla97C06G117020 and ClCHYB may interact to form the stigma color. This study provides a theoretical basis for gene fine mapping and mechanisms for the regulation of stigma color in watermelon.

Key words: watermelon , stigma color , gene mapping , zinc finger protein CONSTANS-LIKE 4

Shuang Pei, Zexu Wu, Ziqiao Ji, Zheng Liu, Zicheng Zhu, Feishi Luan, Shi Liu. 数量性状位点揭示锌指蛋白CONSTANS-LIKE 4与西瓜柱头颜色相关[J]. Journal of Integrative Agriculture, 2024, 23(7): 2292-2305.

Shuang Pei, Zexu Wu, Ziqiao Ji, Zheng Liu, Zicheng Zhu, Feishi Luan, Shi Liu. Quantitative trait loci identification reveals zinc finger protein CONSTANS-LIKE 4 as the key candidate gene of stigma color in watermelon (Citrullus lanatus)[J]. Journal of Integrative Agriculture, 2024, 23(7): 2292-2305.

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数量性状位点揭示锌指蛋白CONSTANS-LIKE 4与西瓜柱头颜色相关

Quantitative trait loci identification reveals zinc finger protein CONSTANS-LIKE 4 as the key candidate gene of stigma color in watermelon (Citrullus lanatus)

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