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作物学报 ›› 2023, Vol. 49 ›› Issue (12): 3239-3249.doi: 10.3724/SP.J.1006.2023.24256

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

基于RNA-seq的甘薯芽变株系类胡萝卜素基因代谢差异分析

赵冬兰*(), 赵凌霄, 刘洋, 张安, 戴习彬, 周志林, 曹清河   

  1. 江苏徐淮地区徐州农业科学研究所 / 农业农村部甘薯生物学与遗传育种重点实验室, 江苏徐州 221131
  • 收稿日期:2022-11-18 接受日期:2023-05-24 出版日期:2023-12-12 网络出版日期:2023-06-14
  • 通讯作者: * 赵冬兰, E-mail: 19991006@jaas.ac.cn* E-mail: 19991006@jaas.ac.cn
  • 基金资助:
    徐州市科技局自然科学基金项目(KC21072);财政部和农业农村部国家现代农业产业技术体系建设专项(Sweetpotato, CARS-10)

Relative expression profile of the related genes with carotenoids metabolism in sweetpotato (Ipomoea batatas) based on RNA-seq data

ZHAO Dong-Lan*(), ZHAO Ling-Xiao, LIU Yang, ZHANG An, DAI Xi-Bin, ZHOU Zhi-Lin, CAO Qing-He   

  1. Xuzhou Institute of Agricultural Sciences in Jiangsu Xuhuai District / Key Laboratory of Biology and Genetic Breeding of Sweetpotato, Ministry of Agriculture and Rural Affairs, Xuzhou 221131, Jiangsu, China
  • Received:2022-11-18 Accepted:2023-05-24 Published:2023-12-12 Published online:2023-06-14
  • Supported by:
    Natural Fund Project of Xuzhou Science and Technology Bureau(KC21072);China Agriculture Research System of MOF and MARA(Sweetpotato, CARS-10)

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摘要：

为研究甘薯类胡萝卜素代谢的分子机制, 本研究以浙薯81及其芽变系为材料, 利用RNA-seq技术研究了浙薯81及其芽变系的薯皮(红色变黄色)和脉基(紫色变绿色)类胡萝卜素代谢相关基因。经过生物信息学分析, 在薯皮和脉基分别筛选出24个和10个差异表达基因(DEGs), 其中有4个DEGs同时存在于薯皮和脉基中。类胡萝卜合成代谢上游途径中, 与浙薯81相比, 芽变系薯皮中的牻牛儿基牻牛儿基焦磷酸合酶基因GGDPS (g29773、g38646)、番茄红素合成酶基因PSY (g11630)和ζ-胡萝卜素异构酶基因Z-ISO (g42608)均显著上调, GGDPS (g29773)仅在突变系中有表达; 脉基部在类胡萝卜素上游代谢途径中没有筛选到DEGs。番茄红素环化分支点后, 在薯皮中检测到2个β-胡萝卜素羟化酶基因CHYB (g33351、g953), 与浙薯81相比, 芽变系中g33351下调表达, g953则上调表达。在薯皮和脉基分别检测到一个玉米黄质环氧化酶基因ZEP, 薯皮中的ZEP (g41700)显著下调, 脉基部的ZEP (g1103)显著上调。类胡萝卜素分解代谢中突变体薯皮中类胡萝卜素裂解双加氧酶基因CCD8 (g21348)下调表达; 2个9-顺式环氧类胡萝卜素加氧酶基因NCED (g30636、g43412)在突变系的薯皮和脉基中均下调表达; 此外在薯皮和脉基分别检测出5个黄氧素脱氢酶基因ABA2, 薯皮中有4个上调表达, 而脉基中的则全部下调表达。利用实时定量PCR (qRT-PCR)验证了部分基因的表达模式, 与RNA-seq分析结果一致。该研究对甘薯类胡萝卜素代谢的分子机制及解析甘薯薯皮和脉基不同颜色突变具有重要参考价值。

关键词: 甘薯, 转录组测序, 类胡萝卜素, 差异基因, 颜色变异

Abstract:

The objective of this study is to explore the molecular mechanism of carotenoid metabolism and color mutation of sweetpotato and to study the carotenoid metabolism related genes by RNA-seq technology. Zheshu 81 and its mutant line with color mutation in root skin (from red to yellow) and leaf vein base (from purple to green) were used as the experimental materials. Bioinformatics analysis showed that the carotenoid biosynthesis (ko00906) pathway was significantly enriched in root skin and leaf vein base. The 24 differentially expressed genes (DEGs) and 10 DEGs in the ko00906 were screened from root skin and vein base, among which four were screened in both two parts, respectively. In the upstream pathway of carotenoid anabolism, compared with Zheshu 81, the geranylgeranyl pyrophosphate synthase gene GGDPS (g29773, g38646), lycopene synthase gene PSY (g11630) and ζ-carotene isomerase gene Z-ISO (g42608) were significantly up-regulated in mutant line, and GGDPS (g29773) was only expressed in mutant lines. DEGs were not screened in the upstream metabolic pathway of carotenoids in vein base. After the branching point of lycopene cyclization, two β-carotene hydroxylase genes CHYB (g33351, g953) were screened in root skin, one up-regulated and the other down-regulated. Two zeaxanthin epoxidase gene ZEP (g41700, g1103) were detected in root skin and vein base, respectively. And g41700 in root skin was significantly down-regulated, and g1103 in vein base was significantly up-regulated. In carotenoid catabolism, CCD8 (g21348), a carotenoid cleavage dioxygenase gene, was significantly up-regulated in root skin of mutant line. Two 9-cis epoxide carotenoid oxygenase genes NCED (g30636, g43412) were down-regulated in root skin and vein base of the mutant line. In addition, ten xanthoxin dehydrogenase ABA2 genes were detected in root skin and vein base, and nine of them were up-regulated. The relative expression levels of some genes by qRT-PCR were highly consistent with those of RNA-seq data. This study provides an important reference for the molecular mechanism of carotenoid metabolism and the analysis of different color mutations in root skin and vein base of sweetpotato.

Key words: sweetpotato, carotenoid, RNA-seq, differentially expressed gene, color variation

图1

浙薯81及其芽变体颜色变异 A, B: 浙薯81和芽变系叶脉基部颜色对比, A为浙薯81, B为芽变系。C: 浙薯81与芽变系薯块膨大初期薯皮对比。D: 浙薯81芽变体薯块。标尺为5 cm。"

图2

浙薯81及其芽变系类胡萝卜素含量"

图3

浙薯81及其突变系差异基因KEGG通路富集散点图 A: 叶脉基部差异基因KEGG富集前20条通路; B: 薯皮差异基因KEGG富集前20条通路。红色方框内是本研究重点关注通路。"

图4

浙薯81及其芽变系类胡萝卜素代谢转录热图 图中基因名称后(LVB)表示该基因是叶脉基部的差异基因, (RS)表示该基因是薯皮的差异基因。"

图5

浙薯81及其突变系薯皮类胡萝卜素相关基因的差异表达 A: 浙薯81及其芽变系薯皮中类胡萝卜素相关DEGs RNA-seq和qRT-PCR表达量的相关性分析; B: 芽变系薯皮中类胡萝卜素相关基因相对于浙薯81的差异表达。*表示在P < 0.05水平差异显著, **表示在P < 0.01水平差异显著。"

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