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Acta Agronomica Sinica ›› 2019, Vol. 45 ›› Issue (8): 1270-1278.doi: 10.3724/SP.J.1006.2019.84129

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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 Online:2019-08-12 Published:2019-04-26
  • Contact: Li-Quan ZHU E-mail:zhuliquan@swu.edu.cn
  • 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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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

Table 1

Information about protein of the PIN-FORMED family in Brassica oleracea"

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

Fig. 1

Phylogenetic tree and gene structure of BoPINs in Brassica oleracea The phylogenetic tree is constructed with MEGA6.0 software on the left and the gene structure map of the BoPINs family on the right."

Fig. 2

Phylogenetic tree of BoPINs gene family in Brassica oleracea"

Fig. 3

Distribution of BoPINs family in Brassica oleracea chromosome"

Fig. 4

Expression patterns of BoPINs gene family after self-pollination and cross-pollination"

Fig. 5

Expression analysis of BoPINs gene family after self-pollination and cross-pollination SI: self-pollination; CP: cross-pollination."

Table 2

Predicted cis-element in the promoter of the BoPINs genes"

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

Fig. 6

Heat-map of members of the BoPINs gene family in different tissues after self-pollination time"

Fig. 7

Changes in mRNA content of auxin biosynthesis key enzyme TAA1 after stigma pollination"

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