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Acta Agronomica Sinica ›› 2020, Vol. 46 ›› Issue (5): 680-689.doi: 10.3724/SP.J.1006.2020.94096

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Genome wide identification and expression analysis of CRK gene family in response to fungal pathogen signals in potato

Wei-Na ZHANG,Yan-Ling FAN,Yi-Chen KANG,Xin-Yu YANG,Ming-Fu SHI,Kai YAO,Zhang-Ping ZHAO,Jun-Lian ZHANG,Shu-Hao QIN()   

  1. College of Horticulture, Gansu Agricultural University, Lanzhou 730070, Gansu, China
  • Received:2019-06-28 Accepted:2019-12-26 Online:2020-05-12 Published:2019-10-11
  • Contact: Shu-Hao QIN E-mail:qinsh@gsau.edu.cn
  • Supported by:
    This study was supported by the Discipline Construction Fund Project of Gansu Agricultural University(GAU-XKJS-2018-225);the National Natural Science Foundation of China(31260311);the Natural Science Foundation of Gansu Province(1606RJZA034);the China Postdoctoral Science Foundation(2012M512042);the China Postdoctoral Science Foundation(2014T70942);the China Agriculture Research System (Potato)(CARS-09-P14)

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Abstract:

Cysteine-rich receptor-like kinase (CRK) plays an important role in plant growth and environmental adaptation. In this study, potato CRK (StCRK) family members were identified, and their physical and chemical characteristics, evolutionary characteristics, subcellular location, chromosome location and expression patterns were analyzed. Eight StCRK members were identified, with amino acid size from 459 to 686 aa, molecular weight of 50.75-77.50 kD, and isoelectric point of 5.84-8.75. StCRKs were mainly located on plasma membrane. CRKs from potato (Solanum tuberosum), apple (Malus pumila Mill.), Thale Cress (Arabidopsis thaliana), rice (Oryza sativa), cotton (G. hirsutum), banana (Musa acuminata), and tomato (Solanum lycopersicum) could be divided into nine subgroups, and StCRKs were belonged to subgroups I (6 members) and VI (2 members). Moreover, StCRKs distributed on chromosomes 2, 3, and 5, contained two tandem repeat gene clusters, including four members. There were many cis-regulated elements in the StCRKs promoter region, which mainly respond to hormones, low temperature, defense and stress signals. After inoculating Phytophthora infestans (Pi) and Fusarium sulphureum (Fs), eight and six StCRKs gene differentially expressed, among them, StCRK4 and StCRK8 had the expression levels by more than eight times. It is speculated that they may respond to multiple fungal signals, and play an important role in potato's broad-spectrum resistance to fungal diseases, and can be used as candidate genes for further needed on disease resistance and functional analysis.

Key words: CRK, cis-elements, bioinformatic analysis, Fusarium sulphureum (Fs), Phytophthora infestans (Pi)

Table 1

Primers of StCRKs"

基因登录号
Gene accession number		 上游引物
Forward primer (5′-3′)		 下游引物
Reverse prime (5′-3′)
AB061263 ATTGGAAACGGATATGCTCCA TCCTTACCTGAACGCCTGTCA
PGSC0003DMG402000515 ACTCTGGCTCTCTACTACAAACAGT CTTCCACACCTGAAACCAAAATGAC
PGSC0003DMG400021394 AAGAGTCCCCTTGTATACAGAACCA ATCGAATTCATACAAGAGGTCCAGC
PGSC0003DMG400013525 CAACACAACCACCATTCCTCAATTC AAGATTGTGTAGAGGAGCTTGGTTG
PGSC0003DMG400018101 CTCCTCCAGATACAAGCAGTTCATC TCACCTGATTGAGAGCTAGAAATGC
PGSC0003DMG400006912 TCGTGTTCAAGAGTTATGTCACCAG AATTCCAAGCAAGTCGAAGTCAGAT
PGSC0003DMG400013524 CATATGTAGTACGCAACCTGAGCAT CTTCAGCATAGCATAGGACACAGTC
PGSC0003DMG400015170 TCATGATCAGACTGTTGACTTCGTC AAGCAGTGAATGGAGCAACAAAATC
PGSC0003DMG400015171 TACAAAAGCCAGTGAAGGAGAAGAC TATAATTGCCTGTTTCTTGAGCGGA

Table 2

CRKs in potato"

基因登录号Gene accession number 基因编号
Genetic code		 氨基酸数
AA size		 分子量
Molecular weight (kD)		 等电点
Isoelectric point		 亚细胞定位
Subcellular localization
Gene ID# STWG 1.0b
PGSC0003DMG402000515 PGSC0003DMP400001023/
PGSC0003DMP400001024		 StCRK1 616 67.40 8.53 PMa, Chlob
PGSC0003DMG400021394 PGSC0003DMP400037082 StCRK2 608 67.23 8.07 PMa, Chlob
PGSC0003DMG400013525 PGSC0003DMP400023929 StCRK3 656 72.73 8.75 PMa, Chlob
PGSC0003DMG400018101 PGSC0003DMP400031539 StCRK4 681 75.52 6.37 PMa, Chlob
PGSC0003DMG400006912 PGSC0003DMP400012233/
PGSC0003DMP400012234/
PGSC0003DMP400012235		 StCRK5 668 75.21 5.84 PMa, Vacub
PGSC0003DMG400013524 PGSC0003DMP400023928/
PGSC0003DMP400023926		 StCRK6 656 73.21 6.42 PMa,b
PGSC0003DMG400015170 PGSC0003DMP400026615/
PGSC0003DMP400026616/
PGSC0003DMP400026617		 StCRK7 459 50.75 7.27 PMa, Cytob
PGSC0003DMG400015171 PGSC0003DMP400026620 StCRK8 686 77.50 7.23 PMa, b

Fig. 1

Phylogenetic tree of CRKs in potato (Solanum tuberosum) (▼), apple (Malus pumila Mill.), Thale Cress (Arabidopsis thaliana), rice (Oryza sativa), cotton (G. hirsutum), banana (Musa acuminata), and tomato (Solanum lycopersicum) "

Fig. 2

Chromosomal distribution of the StCRKs * indicates tandem duplication genes. "

Fig. 3

Gene structure and protein functional domain analysis of StCRKs A: phylogenetic tree of StCRKs; B: gene structure of StCRKs; C: functional domains of StCRKs. "

Fig. 4

Putative regulatory cis-elements in StCRKs promoters A and B: are the abscisic acid responsive elements; C: GARE-motif; D and E: the Gibberellin responsive elements; F and G: the auxin responsive elements; H: the Methyl jasmonate responsive element; I: the Salicylic acid responsive elements; J: the regulatory element for Zein metabolism; K: the Anaerobic responsive elements; L: the anoxic responsive elements; M: the defense and stress responsive elements; N: the flavonoid responsive elements; O: the low-temperature responsive elements. "

Fig. 5

Expression patterns of six StCRKs in response to SA and JA treatments"

Fig. 6

Expression patter of StCRKs in response to Phytophthora infestans (Pi) "

Fig. 7

Expression patter of StCRKs in response to Fusarium sulphureum (Fs) "

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