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作物学报 ›› 2019, Vol. 45 ›› Issue (8): 1270-1278.doi: 10.3724/SP.J.1006.2019.84129

• 研究简报 • 上一篇    下一篇

甘蓝BoPINs家族基因的特征和表达分析

王玉奎1,张贺翠1,白晓璟1,廉小平2,施松梅2,刘倩莹1,左同鸿1,朱利泉1,*()   

  1. 1 西南大学农学与生物科技学院, 重庆 400715
    2 西南大学园艺园林学院, 重庆 400100
  • 收稿日期:2018-10-15 接受日期:2019-04-15 出版日期:2019-08-12 网络出版日期:2019-04-26
  • 通讯作者: 朱利泉
  • 作者简介:E-mail: wangyuk0808@163.com
  • 基金资助:
    本研究由国家自然科学基金项目(31572127);中央高校基本科研业务费项目资助(XDJK2017C023)

Characteristics and expression analysis of BoPINs family genes in Brassica oleracea

WANG Yu-Kui1,ZHANG He-Cui1,BAI Xiao-Jing1,LIAN Xiao-Ping2,SHI Song-Mei2,LIU Qian-Ying1,ZUO Tong-Hong1,ZHU Li-Quan1,*()   

  1. 1 College of Agriculture and Biotechnology, Southwest University, Chongqing 400715, China
    2 College of Horticulture and Gardening, Southwest University, Chongqing 400100, China
  • Received:2018-10-15 Accepted:2019-04-15 Published:2019-08-12 Published online:2019-04-26
  • Contact: Li-Quan ZHU
  • Supported by:
    This study was supported by the National Natural Science Foundation of China(31572127);the Basic Research Fund of the Central University(XDJK2017C023)

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摘要：

为了探索植物生长素极性运输载体蛋白编码基因BoPINs家族参与甘蓝自交不亲和性的成员数目与参与方式, 本文通过转录组分析获得BoPINs家族在甘蓝自花和异花授粉后的表达情况, 利用分子生物学技术和生物信息学方法对该家族的基因结构、蛋白进化亲缘关系和表达模式等特征进行分析。结果表明, 甘蓝BoPINs基因家族包含8个成员, 含有5波浪线9个外显子和4~8个内含子; 其编码的蛋白质的氨基酸残基数在350波浪线650之间, 相对分子质量为38波浪线70 kD; 除了BoPIN5和BoPIN8不含中间亲水区以外, 其余6个BoPINs家族成员都含有位于两端的疏水区和中间亲水环, 它们可能以膜锚定蛋白的形式发挥作用; 甘蓝BoPINs与芜菁BrPINs、拟南芥AtPINs基因家族亲缘关系较近; 染色体定位分析表明, BoPIN1-1、BoPIN3-1、BoPIN3-2和BoPIN6与S位点之间发生不同程度的连锁; 启动子活性分析表明, BoPINs家族蛋白参与甘蓝SI反应, 可能受IAA、ABA等激素相互交叉影响; BoPIN1-1、BoPIN1-2、BoPIN2、BoPIN3-1、BoPIN3-2、BoPIN4、BoPIN6、BoPIN7-1和BoPIN7-2在柱头中表达量均较高; 数据表达谱和荧光定量分析表明, 8个家族成员中的6个BoPINs基因在自花授粉后下调表达; 自花授粉后柱头生长素含量降低, 与SI反应呈负相关。因此, 在甘蓝BoPINs家族的8个成员中有6个BoPINs基因家族成员可能在膜上以负调节方式调控自交不亲和反应。

关键词: 甘蓝, 生长素, 自花授粉, BoPINs家族, 自交不亲和

Abstract:

In order to explore the number and expression of the BoPINs gene family participating in self-incompatibilty of Brassica, their expression after self-pollination and cross-pollination were detected by transcriptome analysis, and the corresponding gene structure, phylogenetic tree and expression patterns of the family were further analyzed by bioinformatics. This gene family contained 5-9 exons and 4-8 introns. The amino acid of the encoding protein residues were between 350 and 650 and had molecular weights ranging from 38 kD to 70 kD. Except that BoPIN5 and BoPIN8 did not contain internal hydrophilic cytoplasmic regions, the remaining six BoPINs proteins contained a hydrophobic region at both ends and an internal hydrophilic ring, showing they located on membrane. The evolutionary analysis indicated that BoPINs were closely related to the BrPINs and the AtPINs gene family. Chromosome localization analysis indicated that BoPIN1-1, BoPIN3-1, BoPIN3-2, and BoPIN6 members of the family were linked to S-loucs to different degrees. Tissue-specific expression analysis indicated that BoPIN1-1, BoPIN1-2, BoPIN2, BoPIN3-1, BoPIN3-2, BoPIN4, BoPIN6, BoPIN7-1, and BoPIN7-2 had higher expression levels in the stigma. Data expression profiling and fluorescence quantitative analysis indicated that six of the eight BoPINs genes were down-regulated after self-pollination. All these results indicate that six members of the eight BoPIN gene family members on the membrane may participate in the self-incompatibility response of Brassica oleracea in a negative regulatory manner.

Key words: Brassica oleracea, auxin, self-pollination, BoPINs family, self-incompatibility

表1

甘蓝BoPINs家族蛋白信息"

蛋白名称
Protein name		 转录组号
Transcriptome No.		 染色体定位
Chromosome localization		 氨基酸
长度
Amino acid length		 等电点
Isoelectric point (pI)		 相对分子量
Molecular weight (kD)		 平均亲水指数
Average hydropathic index		 不稳定系数
Instability coefficient		 亚细胞定位
Subcellular localization
BoPIN1-1 Bo2g080660 C6 617 9.2 66.50 0.121 39.70 Cell membrane
BoPIN1-2 Bo6g116850 C6 619 9.1 66.91 0.075 43.50 Cell membrane
BoPIN2 Bo3g019460 C3 652 9.2 70.46 0.058 40.66 Cell membrane
BoPIN3-1 Bo6g094450 C6 642 8.6 69.63 0.138 40.11 Cell membrane
BoPIN3-2 Bo6g112590 C6 635 8.2 68.40 0.184 40.11 Cell membrane
BoPIN4 Bo2g134160 C2 599 7.2 65.03 0.265 36.77 Cell membrane
BoPIN5 Bo9g165420 C9 362 9.4 39.71 0.702 44.60 Endoplasmic reticulum
BoPIN6 Bo6g121080 C6 567 9.2 61.69 0.409 37.67 Endoplasmic reticulum
BoPIN7-1 Bo6g112720 C6 577 8.5 63.09 0.272 39.44 Cell membrane
BoPIN7-2 Bo7g056420 C7 603 6.8 65.47 0.239 31.78 Cell membrane
BoPIN8 Bo9g163100 C9 348 6.2 38.22 0.703 36.56 Endoplasmic reticulum

图1

甘蓝BoPINs的进化树和基因结构图 左侧为进化树, 用MEGA6.0软件构建; 右侧为甘蓝BoPINs家族的基因结构图。"

图2

甘蓝家族BoPINs的系统进化树"

图3

BoPINs家族在甘蓝染色体的分布"

图4

BoPINs基因家族在自花授粉和异花授粉后的表达模式"

图5

甘蓝BoPINs家族在自花和异花授粉后的表达分析"

表2

BoPINs基因启动子的顺式作用元件"

基因
Gene		 生长素
响应元件
Auxin response element		 水杨酸
响应元件
Salicylic acid response element		 脱落酸
响应元件
Abscisic acid response element		 胁迫响应元件
Stress
response
element		 赤霉素
响应元件
Gibberellin response element		 厌氧诱导元件
Anaerobic induction
element		 光响应元件
Light responsive element		 茉莉酸甲酯
响应元件
MeJA response element
BoPIN1-1 — 2 2 1 1 1 7 1
BoPIN1-2 — 1 1 2 1 1 6 2
BoPIN2 — 1 1 1 — 1 8 2
BoPIN3-1 1 2 — 2 — 1 5 —
BoPIN3-2 1 2 — 1 — 2 9 —
BoPIN4 — 2 1 2 12 12 2
BoPIN5 — 1 — 3 1 1 5 2
BoPIN6 — 4 — — 1 1 9 1
BoPIN7-1 1 2 1 1 — — 14 —
BoPIN7-2 1 4 1 2 — — 2 1
BoPIN8 2 1 1 1 — 1 9 2

图6

BoPINs基因家族成员在自花授粉后不同组织中的表达量分析"

图7

柱头授粉后生长素生物合成关键酶TAA1的mRNA含量变化"

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