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作物学报 ›› 2022, Vol. 48 ›› Issue (1): 249-258.doi: 10.3724/SP.J.1006.2022.14019

• 研究简报 • 上一篇    下一篇

马铃薯CrRLK1Ls基因家族的鉴定及响应晚疫病菌信号的表达分析

余慧芳(), 张卫娜, 康益晨, 范艳玲, 杨昕宇, 石铭福, 张茹艳, 张俊莲, 秦舒浩*()   

  1. 甘肃农业大学园艺学院, 甘肃兰州 730070
  • 收稿日期:2021-01-31 接受日期:2021-06-18 出版日期:2022-01-12 网络出版日期:2021-07-23
  • 通讯作者: 秦舒浩
  • 作者简介:E-mail: 2080277369@qq.com
  • 基金资助:
    国家自然科学基金项目(32060441);国家现代农业产业技术体系建设专项(CARS-09-P14);甘肃农业大学园艺学科建设基金项目资助(GAU-XKJS-2018-225)

Genome-wide identification and expression patterns in response to signals from Phytophthora infestans of CrRLK1Ls gene family in potato

YU Hui-Fang(), ZHANG Wei-Na, KANG Yi-Chen, FAN Yan-Ling, YANG Xin-Yu, SHI Ming-Fu, ZHANG Ru-Yan, ZHANG Jun-Lian, QIN Shu-Hao*()   

  1. College of Horticulture, Gansu Agricultural University, Lanzhou 730070, Gansu, China
  • Received:2021-01-31 Accepted:2021-06-18 Published:2022-01-12 Published online:2021-07-23
  • Contact: QIN Shu-Hao
  • Supported by:
    National Natural Science Foundation of China(32060441);China Agriculture Research System(CARS-09-P14);Discipline Construction Fund Project of Gansu Agricultural University(GAU-XKJS-2018-225)

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摘要：

Catharanthus roseus receptor-like kinase1-like kinases (CrRLK1Ls)是植物所特有的一类蛋白类受体激酶, 在维持细胞壁完整性、细胞间通讯、生物和非生物应激反应中发挥重要作用。本研究鉴定了马铃薯CrRLK1Ls (StCrRLK1Ls)基因家族的成员数量, 并对其理化特性、染色体位置、进化特征、亚细胞定位和对晚疫病菌侵染时的表达模式进行全面分析。结果显示, 共获得17个StCrRLK1Ls基因, 其氨基酸序列大小为753~997 aa, 分子量和等电点分别为83.34~108.69 kD和5.30~7.56, 主要位于质膜。进化分析将马铃薯、拟南芥、水稻、苹果和番茄的CrRLK1Ls家族成员分为7个亚组, 马铃薯CrRLK1Ls家族成员分布于亚组III、IV、V、VI和VII。StCrRLK1Ls不均匀的分布于8条染色体上, 存在3个串联重复基因簇, 包含6个基因。此外, StCrRLK1Ls启动子区域存在响应植物激素、防卫和逆境等多种顺式调控元件。‘大西洋’和‘陇薯7号’接种晚疫病菌(Phytophthora infestans, Pi)后, 大量StCrRLK1Ls为差异表达。其中, StCrRLK1L8和StCrRLK1L10的相对表达量在晚疫病菌侵染期间明显上调; 在‘大西洋’接种Pi 8 d时, StCrRLK1L11的表达量是对照的9.74倍, 推断其可能在马铃薯响应病原真菌的过程中起重要的作用, 可作为进一步开展抗病研究和功能分析的候选基因。

关键词: CrRLK1Ls, 生物信息学分析, 马铃薯晚疫病菌, 表达模式

Abstract:

Catharanthus roseus receptor-like kinase1-like kinases (CrRLK1Ls), a class of plant-specific protein receptor kinases, play an important role in maintaining cell wall integrity, cell-to-cell communications, and biotic and abiotic stress response. In this study, StCrRLK1Ls family members were identified, and their physical and chemical characteristics, chromosome location, evolutionary characteristics, subcellular location, and the expression patterns were comprehensively analyzed. The results showed that 17 StCrRLK1Ls members were detected, with amino acid size (753-997 aa), molecular weight (83.34-108.69 kD), and isoelectric point (5.30-7.56). Among them, StCrRLK1Ls were mainly predicted on plasma membrane. Evolutionary analysis indicated that CrRLK1Ls from potato, rice, apple, Arabidopsis, and tomato could be divided into seven subgroups, and StCrRLK1Ls were distributed in subgroups III, IV, V, VI, and VII. Moreover, StCrRLK1Ls were distributed on eight chromosomes, and there are three tandem repeat gene clusters, including six members. In addition, there were many cis-elements in the promoter region of StCrRLK1Ls, which mainly responded to hormones, defense, and stress signals. After the inoculation with Phytophthora infestans (Pi), a large number of StCrRLK1Ls were differentially expressed in Atlantic and Longshu 7, and the relative expression levels of StCrRLK1L8 and StCrRLK1L10 were obviously up-regulated during Pi infection. The relative expression levels of StCrRLK1L11 were 9.74 times compared with the control at eight days Atlantic inoculated Pi. We speculated that they could be used as the candidate genes for disease resistance and further functional analysis, which might play an important role in response to pathogenic fungi of potato.

Key words: CrRLK1Ls, bioinformatic analysis, Phytophthora infestans (Pi), the relative expression pattern

表1

马铃薯CrRLK1Ls基因特性"

基因登录号
Gene ID		 基因名称
Gene name		 氨基酸数
Amino acid number (aa)		 分子量
Molecular
Weight (kD)		 等电点
Isoelectric
point		 不稳定
性系数
Instability
index (II)		 亲水性
指数
GRAVY		 脂溶
指数
Aliphatic
index		 亚细胞定位
Subcellular localization
PGSC0003DMP400037841 StCrRLK1L1 854 95.05 6.39 41.44 -0.282 77.88 PMa, ECb
PGSC0003DMP400065940 StCrRLK1L2 873 96.03 5.67 42.18 -0.207 79.34 PMa,b
PGSC0003DMP400036732 StCrRLK1L3 868 96.30 5.57 37.21 -0.274 83.13 PMa, ECb
PGSC0003DMP400002467 StCrRLK1L4 856 95.43 6.13 42.26 -0.202 85.07 PMa,b
PGSC0003DMP400002464 StCrRLK1L5 873 97.31 6.25 39.14 -0.256 81.41 PMa,b
PGSC0003DMP400026188 StCrRLK1L6 818 91.57 5.80 33.92 -0.200 85.38 PMa,b
PGSC0003DMP400045697 StCrRLK1L7 818 91.27 7.56 31.77 -0.083 89.44 PMa,b
PGSC0003DMP400042695 StCrRLK1L8 753 83.34 6.11 41.43 -0.122 86.33 PMa, Chlob
PGSC0003DMP400040508 StCrRLK1L9 842 92.84 6.59 37.71 0.010 92.46 PMa,b
PGSC0003DMP400040693 StCrRLK1L10 818 91.32 5.55 32.47 -0.068 92.19 PMa,b
PGSC0003DMP400012927 StCrRLK1L11 997 108.69 7.10 36.74 -0.213 84.40 PMa,b
PGSC0003DMP400012941 StCrRLK1L12 889 97.92 5.51 38.65 -0.275 82.35 PM, Chlo
PGSC0003DMP400017117 StCrRLK1L13 829 90.65 5.77 36.39 -0.051 85.68 PMa,b
PGSC0003DMP400052063 StCrRLK1L14 889 97.45 5.93 39.97 -0.211 80.37 PMa,b
PGSC0003DMP400043439 StCrRLK1L15 834 91.29 5.30 37.55 0.018 94.46 PMa,b
PGSC0003DMP400006076 StCrRLK1L16 872 96.70 5.96 38.47 -0.190 81.08 PMa,b
PGSC0003DMP400027254 StCrRLK1L17 834 91.51 5.73 35.37 -0.115 83.78 PMa,b

图1

马铃薯、拟南芥、水稻、番茄和苹果CrRLK1Ls家族进化分析树"

图2

马铃薯CrRLK1Ls染色体定位"

图3

马铃薯CrRLK1Ls基因家族的进化树、基因结构和Motif分析 A: 马铃薯CrRLK1Ls基因家族进化树; B: 马铃薯CrRLK1Ls基因家族结构分析; C: 马铃薯CrRLK1Ls基因家族Motif分析。"

图4

马铃薯CrRLK1Ls启动子区域响应植物生长调节剂和逆境的顺式调控元件 A: ABRE是脱落酸响应元件; B: CGTCA-motif 和TGACG-motif是茉莉酸甲酯响应元件; C: TCA-element是水杨酸响应元件; D: P-box和TATC-box是赤霉素响应元件; E: TGA-element和AuxRR-core是生长素响应元件; F: TC-rich repeats是防御和压力响应元件; G: MBS是干旱响应元件; H: LTR是低温响应元件; I: MBSI是黄酮类代谢响应元件; J: AT-rich sequence是激活响应元件; K: ciradian是昼夜节律响应元件; L: ARE是厌氧诱导响应元件; M: O2-site是玉米醇溶蛋白代谢调控元件。"

图5

ABA和GA3处理后6个马铃薯CrRLK1Ls基因的表达情况 *、**分别表示处理与对照在0.05和0.01水平上显著差异。"

图6

马铃薯CrRLK1Ls响应晚疫病菌侵染的表达模式 *和**分别表示处理与对照在0.05和0.01水平上显著差异。"

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