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作物学报

作物学报 ›› 2023, Vol. 49 ›› Issue (12): 3250-3260.doi: 10.3724/SP.J.1006.2023.34002

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

木薯花性别分化关键时期的转录组分析及雌花分化相关候选基因的筛选

陈会鲜(), 梁振华, 黄珍玲, 韦婉羚, 张秀芬, 杨海霞, 李恒锐*(), 何文*(), 李天元, 兰秀, 阮丽霞, 蔡兆琴, 农君鑫   

  1. 广西南亚热带农业科学研究所, 广西龙州 532415
  • 收稿日期:2023-01-05 接受日期:2023-05-24 出版日期:2023-12-12 网络出版日期:2023-06-06
  • 通讯作者: * 李恒锐, E-mail: lihengrui88@163.com; 何文, E-mail: 675559090@qq.com
  • 作者简介:E-mail: 798555436@qq.com
  • 基金资助:
    国家重点研发计划项目(2020YFD1000603-10);广西青年科学基金项目(2023GXNSFBA026150);广西农业科学院科技发展基金项目(Guinongke 2022JM96);崇左市科技计划项目(Chongkegong 2021ZC18);广西农业科学院基本科研业务专项(Guinongke 2023YM44)

Transcriptomic profile of key stages of sex differentiation in cassava flowers and discovery of candidate genes related to female flower differentiation

CHEN Hui-Xian(), LIANG Zhen-Hua, HUANG Zhen-Ling, WEI Wan-Ling, ZHANG Xiu-Fen, YANG Hai-Xia, LI Heng-Rui*(), HE Wen*(), LI Tian-Yuan, LAN Xiu, RUAN Li-Xia, CAI Zhao-Qin, NONG Jun-Xin   

  1. Guangxi Institute of Sub-tropical Agricultural Sciences, Longzhou 532415, Guangxi, China
  • Received:2023-01-05 Accepted:2023-05-24 Published:2023-12-12 Published online:2023-06-06
  • Contact: * E-mail: lihengrui88@163.com; E-mail: 675559090@qq.com
  • Supported by:
    National Key Research and Development Program of China(2020YFD1000603-10);Guangxi Youth Science Foundation Project(2023GXNSFBA026150);Guangxi Academy of Agricultural Sciences Development Fund Project(Guinongke 2022JM96);Science and Technology Plan Project of Special Project for Basic Scientific Research of Chongzuo(Chongkegong 2021ZC18);Guangxi Academy of Agricultural Sciences(Guinongke 2023YM44)

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摘要：

针对木薯雌花数量严重缺乏的育种难题, 本试验以木薯品种‘新选048’为试材, 测定木薯花性别分化关键时期雌花和雄花的转录组数据, 分析差异表达基因的功能及其可能参与的调控通路, 揭示木薯雌花分化的分子机制, 挖掘木薯雌花分化的相关候选基因, 并采用qRT-PCR方法对测序结果进行验证。结果表明: 在木薯花性别分化关键时期, 雌花和雄花共有545个差异基因, 其中48.63%的差异基因富集到花器官的形态建成及发育的GO途径, 且富集到植物苯丙素生物合成、植物激素信号传导2个代谢通路的基因最多, 4个花性别分化基因AGL11、YABBY4、CRC、SUP在雌花中显著上调表达, 4个细胞分裂素信号通路基因HK4、HPt4、ARR8、ARR12在雌花中显著上调表达, 生长素的早期响应基因IAA14、GH3、SAUR22、SAURR20在雌花中显著下调表达。因此, 木薯花性别分化主要涉及花粉、配子体、花器官的形态建成及发育的生物途径和植物苯丙素生物合成、植物激素信号传导2个代谢通路, 细胞分裂素和生长素可能参与了木薯雌花分化过程, AGL11、YABBY4、CRC、SUP、HK4、HPt4、ARR8、ARR12可能是木薯雌花分化的正调控基因。

关键词: 木薯, 分化关键时期, 转录组, 雌花, 基因

Abstract:

To solve the breeding problem of the serious shortage of female cassava flowers, cassava variety ‘Xinxianxuan 048’ was used as the experimental material. Transcriptional sequencing technology was used to analyze the biological information of differentially expressed genes in female and male flowers during the critical period of cassava flower sex differentiation, explore the functions of differentially expressed genes and possible regulatory pathways involved, excavate candidate genes related to female differentiation, and verify the sequencing results by qRT-PCR method. The results showed that: There were 545 differentially expressed genes between male and female cassava flowers at the critical stage of gender differentiation. Among them, 48.63% of the differential genes were enriched in GO pathway of floral organ morphogenesis and development, and the genes enriched in plant phenylpropanol biosynthesis and plant hormone signal transduction were the most. AGL11, YABBY4, CRC, SUP, and other flower sex differentiation genes were significantly up-regulated in female flowers. Four genes of the cytokinin signaling pathway, including HK4, HPt4, ARR8, and ARR12, were significantly up-regulated in female flowers, while IAA14, GH3, SAUR22, and SAURR20, the early auxin response genes, were significantly down-regulated in female flowers. Therefore, the sexual differentiation of cassava flowers mainly involved the biological pathways of pollen, gametophyte, floral organ morphogenesis and development, and two metabolic pathways of plant phenylpropanoid biosynthesis and plant hormone signal transduction. Cytokinin and auxin might be the main hormones in cassava female flower differentiation. AGL11, YABBY4, CRC, SUP, HK4, HPt4, ARR8, and ARR12 may be positive regulators of cassava female flower differentiation.

Key words: cassava, key stages of sex differentiation, transcriptome, female flowers, gene

表1

荧光定量PCR引物设计"

基因名称
Gene name		 正向引物
Forward primer (5°-3°)		 反向引物
Reverse primer (5°-3°)		 基因登录号
Gene ID
AGL11 TATGAATTGTCAGTCCTTTGTG TCTAACAGGCATCTAATGGGT LOC110616214
CRC CACAGTTCTTGCGGTTGG TTGGCCTTGAAAAGGAGGT LOC110615357
YABBY4 CAGATTTGCTACGTCCAATGTG AATGGCACGGTTCACTGGG LOC110627391
MADS9 CACCTGAAAGGGGAGGAT TGACGAACACTGGCAAGG LOC110607164
TM6 TTAGCCCTACCATAACGA ATAATGCGTGCTCCACAG LOC110630551
ARR12 TGCCGAGGTTGAGAAATC TGCTCCTGCCGTTTGATC LOC110631158
HPt4 GCAGGGCAGGAAATGGAG CTTGGGGCGACATGCAGT LOC110627048
HK4 GCGGAGTAGCCTATGCAC TTCCCGGTAGCTCTAGCC LOC110603154
TGA9 CTTCACCAACTTCGCCGGATAC AGGACGGGATGCCCAGAGG LOC110626841
TGA10 GCTTATGACTTGGGAGAA CATGGACGGCTGATTTGA LOC110620640

表2

碱基信息统计表"

样本
Sample		 干净序列读数(个)
Read number		 碱基数
Base number (G)		 Q20 (%) Q30 (%)
A1-1 50,981,844 7.25 95.680 87.990
A1-2 35,421,366 4.83 97.975 93.795
A1-3 39,490,380 5.40 97.780 93.375
A2-1 33,138,892 4.48 97.765 93.350
A2-2 31,980,834 4.38 96.455 89.915
A2-3 42,204,558 5.79 98.130 94.145

图1

差异基因的统计图(雌花)"

图2

差异基因的维恩图图"

图3

雄花 vs 雌花的火山图 红色区域表示雄花上调, 蓝色区域表示雄花下调基因。"

图4

前30条显著富集到的GO条目"

图5

差异基因显著富集的GO条目"

图6

前20条显著富集到的KEEG通路"

图7

花性别分化相关候选基因热图 颜色代表响应的的FPKM值, 数值越大, 颜色越红, 从左至右样本依次为雌花-1 (A1-1)、雌花-2 (A1-2)、雌花-3 (A1-3)、雄花-1 (A2-1)、雄花-2 (A2-2)、雄花-3 (A2-3)。"

图8

细胞分裂素信号通路基因表达模式分析 颜色代表响应的的FPKM值, 从左至右样本标号参照图7。"

图9

生长素响应基因的表达模式分析 颜色代表响应的的FPKM值, 从左至右样本标号参照图7。"

图10

差异表达基因qRT-PCR验证的相对表达量 柱状图上小写字母表示雌花、雄花之间10个候选基因的表达量差异达显著水平(P < 0.05)。"

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