Preliminary Analysis of CsCalS5 and Callose Deposition in Citrus sinensis Infected with Candidatus Liberibacter asiaticus

doi:10.16420/j.issn.0513-353x.2020-0282

Acta Horticulturae Sinica ›› 2021, Vol. 48 ›› Issue (2): 276-288.doi: 10.16420/j.issn.0513-353x.2020-0282

• Research Papers • Previous Articles Next Articles

Preliminary Analysis of CsCalS5 and Callose Deposition in Citrus sinensis Infected with Candidatus Liberibacter asiaticus

ZHANG Qingwen, QI Jingjing, XIE Yu, XIE Zhu, PENG Yun, LI Qiang, PENG Aihong, ZOU Xiuping, HE Yongrui, CHEN Shanchun^*(), YAO Lixiao^*()

National Center for CitrusVarieties Improvement,Chongqing 400712,China

Received:2020-06-24 Revised:2020-09-07 Online:2021-02-25 Published:2021-03-09
Contact: CHEN Shanchun,YAO Lixiao E-mail:chenshanchun@cric.cn;yaolixiao@cric.cn

Abstract

Abstract:

Callose synthase is a key enzyme to control callose synthesis and plays an important role in plant growth and stress resistance. The CsCalS5 gene and promoter sequence of Citrus sinensis‘Jincheng’ were cloned and analyzed. Its tissue expression and induced expression with exogenous phytohormones,Candidatus Liberibacter asiaticus（CLas）and Xanthomonas citri subsp. Citri（Xcc）were detected by real-time fluorescent quantitative PCR. The callose deposition in vein of CLas-infected or Xcc-infected C. sinensis was observed by tissue section staining. The CsCalS5 promoter sequence contained abscisic acid （ABA）and pathogen response elements. The CsCalS5 gene encoded a transmembrane protein composed of 1 952 amino acids containing the conserved domains FKS1 and β-1,3-glucan synthetase functional domain. CsCalS5 was highly expressed in the stem of C. sinensis,and could be induced by ABA. The expression of CsCalS5 in CLas-infected leaves was 4.02 times of that in the healthy control,and the callose was obviously deposited in the phloem of vein of CLas-infected leaves. After Xcc infection,callose deposition was not significantly increased in the veins phloem,the expression of CsCalS5 was not significantly different from healthy controls. In conclusion,CLas might promote the deposition of callose in the phloem by up-regulating the expression of CsCalS5 in C. sinensis,which could be regulated by ABA.

Key words: Citrus sinensis, callose synthase, CsCalS5, expression analysis, Huanglongbing

CLC Number:

S666

ZHANG Qingwen, QI Jingjing, XIE Yu, XIE Zhu, PENG Yun, LI Qiang, PENG Aihong, ZOU Xiuping, HE Yongrui, CHEN Shanchun, YAO Lixiao. Preliminary Analysis of CsCalS5 and Callose Deposition in Citrus sinensis Infected with Candidatus Liberibacter asiaticus[J]. Acta Horticulturae Sinica, 2021, 48(2): 276-288.

Figures/Tables 10

References 51

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引物名称 Primer name	引物序列（5′-3′） Primer sequence
CsCalS5-1	F：ATAGAGGCAAAAGATGTCACAGAGA;R：TTTTGCCGATCCATCTGTCAC
CsCalS5-2	F：CGTGCTGACGCTGATTTCT;R：AATAACCTCTTTCTCTCGTTCTCC
CsCalS5-3	F：GCTAGCAAGATACCCATAGCATT;R：CTTAAGGAATTCTTGAAGCAAATTC
CsCalS5-4	F：GGAAACCTGAAAATCAAAACCA;R：TCACTCCTTACTTTTGGAAGACCT
CsCalS5P	F：CTCATGCTTTATGCCTTT;R：CTTTTGCCTCTATAAATATGAT
Actin	F：CATCCCTCAGCACCTTCC;R：CCAACCTTAGCACTTCTCC
qCsCalS5	F：AACGTTCTGGTGATGCTCGT;R：AGCCTTCGCAAGAGCTGAAT
OI1/OI2C	F：GCGCGTATGCAATACGAGCGGCA;R：GCCTCGCGACTTCGCAACCCAT

引物名称 Primer name	引物序列（5′-3′） Primer sequence
CsCalS5-1	F：ATAGAGGCAAAAGATGTCACAGAGA;R：TTTTGCCGATCCATCTGTCAC
CsCalS5-2	F：CGTGCTGACGCTGATTTCT;R：AATAACCTCTTTCTCTCGTTCTCC
CsCalS5-3	F：GCTAGCAAGATACCCATAGCATT;R：CTTAAGGAATTCTTGAAGCAAATTC
CsCalS5-4	F：GGAAACCTGAAAATCAAAACCA;R：TCACTCCTTACTTTTGGAAGACCT
CsCalS5P	F：CTCATGCTTTATGCCTTT;R：CTTTTGCCTCTATAAATATGAT
Actin	F：CATCCCTCAGCACCTTCC;R：CCAACCTTAGCACTTCTCC
qCsCalS5	F：AACGTTCTGGTGATGCTCGT;R：AGCCTTCGCAAGAGCTGAAT
OI1/OI2C	F：GCGCGTATGCAATACGAGCGGCA;R：GCCTCGCGACTTCGCAACCCAT

元件名称 Element name	序列 Sequence	数量 Number	功能 Function	位置 Location
GT1 box	GAAAAA	2	病原菌和盐响应 Pathogen or salt-induced responsiveness	-264（+）/-976（-）
dof box	AAAG	23	病原菌响应Pathogen responsiveness	-1281/-1178/-1097/-974/-927/ -838/-260/-161/-148/-81/-7（+）-1486/-1340/-1264/-1060/-869/-707/-656/-610/-545/-479/-207/-85（-）
W box	TGACT	2	病原菌和损伤响应 Pathogen and wound responsiveness	-474（+）/-582（+）
GARE-motif	TCTGTTG	2	赤霉素响应元件Gibberellin-responsive element	-281（+）
	CCTTTT	2	赤霉素响应元件Gibberellin-responsive element	-1490（+）
TCA-element	CCATCTTTTT	1	水杨酸响应元件Salicylic acid responsiveness	-1067（+）
ABRE	ACGTG	1	脱落酸响应元件Abscisic acid responsiveness	-814（+）
WUN-motif	AAATTACTA	1	损伤响应元件Wound responsiveness	-1369（-）
MYC	CATTTG	2	干旱和冻害响应元件 Drought and freeze responsiveness	-1473（+）/-1400（-）
MBS	CAACTG	1	参与干旱诱导Involved in drought-inducibility	-1259（-）
ARE	AAACCA	2	厌氧诱导所需Essential for the anaerobic induction	-1255（+）/-456（-）
MBSI	TTTTTACGGTTA	1	参与类黄酮合成基因的调控 Involved in flavonoid biosynthetic genes regulation	-131（+）
RY-element	CATGCATG	1	参与种子特异性调控 Involved in seed-specific regulation	-696（-）
GATA-motif	AAGATAAGATT	1	光响应元件Light responsiveness	-1400（-）
GT1-motif	GGTTAA	1	光响应元件Light responsiveness	-1336（-）
chs-CMA2a	TCACTTGA	1	光响应元件Light responsiveness	-1268（-）
Box 4	ATTAAT	1	光响应元件Light responsiveness	-1293（+）
LAMP-element	CTTTATCA	1	光响应元件Light responsiveness	-1123（+）
G-box	TACGTG	2	光响应元件Light responsiveness	-810（+）
G-box	GGTTAAT	2	光响应元件Light responsiveness	-194（+）

元件名称 Element name	序列 Sequence	数量 Number	功能 Function	位置 Location
GT1 box	GAAAAA	2	病原菌和盐响应 Pathogen or salt-induced responsiveness	-264（+）/-976（-）
dof box	AAAG	23	病原菌响应Pathogen responsiveness	-1281/-1178/-1097/-974/-927/ -838/-260/-161/-148/-81/-7（+）-1486/-1340/-1264/-1060/-869/-707/-656/-610/-545/-479/-207/-85（-）
W box	TGACT	2	病原菌和损伤响应 Pathogen and wound responsiveness	-474（+）/-582（+）
GARE-motif	TCTGTTG	2	赤霉素响应元件Gibberellin-responsive element	-281（+）
	CCTTTT	2	赤霉素响应元件Gibberellin-responsive element	-1490（+）
TCA-element	CCATCTTTTT	1	水杨酸响应元件Salicylic acid responsiveness	-1067（+）
ABRE	ACGTG	1	脱落酸响应元件Abscisic acid responsiveness	-814（+）
WUN-motif	AAATTACTA	1	损伤响应元件Wound responsiveness	-1369（-）
MYC	CATTTG	2	干旱和冻害响应元件 Drought and freeze responsiveness	-1473（+）/-1400（-）
MBS	CAACTG	1	参与干旱诱导Involved in drought-inducibility	-1259（-）
ARE	AAACCA	2	厌氧诱导所需Essential for the anaerobic induction	-1255（+）/-456（-）
MBSI	TTTTTACGGTTA	1	参与类黄酮合成基因的调控 Involved in flavonoid biosynthetic genes regulation	-131（+）
RY-element	CATGCATG	1	参与种子特异性调控 Involved in seed-specific regulation	-696（-）
GATA-motif	AAGATAAGATT	1	光响应元件Light responsiveness	-1400（-）
GT1-motif	GGTTAA	1	光响应元件Light responsiveness	-1336（-）
chs-CMA2a	TCACTTGA	1	光响应元件Light responsiveness	-1268（-）
Box 4	ATTAAT	1	光响应元件Light responsiveness	-1293（+）
LAMP-element	CTTTATCA	1	光响应元件Light responsiveness	-1123（+）
G-box	TACGTG	2	光响应元件Light responsiveness	-810（+）
G-box	GGTTAAT	2	光响应元件Light responsiveness	-194（+）

Preliminary Analysis of CsCalS5 and Callose Deposition in Citrus sinensis Infected with Candidatus Liberibacter asiaticus

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[2]	LI Zhenxi, PAN Ruixuan, XU Meirong, ZHENG Zheng, DENG Xiaoling. Development of Duplex Real-time PCR Assay of‘Candidatus Liberibacter asiatics’ [J]. Acta Horticulturae Sinica, 2023, 50(1): 188-196.
[3]	GAO Yanlong, WU Yuxia, ZHANG Zhongxing, WANG Shuangcheng, ZHANG Rui, ZHANG De, WANG Yanxiu. Bioinformatics Analysis of Apple ELO Gene Family and Its Expression Analysis Under Low Temperature Stress [J]. Acta Horticulturae Sinica, 2022, 49(8): 1621-1636.
[4]	QIU Ziwen, LIU Linmin, LIN Yongsheng, LIN Xiaojie, LI Yongyu, WU Shaohua, YANG Chao. Cloning and Functional Analysis of the MbEGS Gene from Melaleuca bracteata [J]. Acta Horticulturae Sinica, 2022, 49(8): 1747-1760.
[5]	ZHENG Lin, WANG Shuai, LIU Yunuo, DU Meixia, PENG Aihong, HE Yongrui, CHEN Shanchun, ZOU Xiuping. Gene Cloning and Expression Analysis of NAC Gene in Citrus in Response to Huanglongbing [J]. Acta Horticulturae Sinica, 2022, 49(7): 1441-1457.
[6]	MA Weifeng, LI Yanmei, MA Zonghuan, CHEN Baihong, MAO Juan. Identification of Apple POD Gene Family and Functional Analysis of MdPOD15 Gene [J]. Acta Horticulturae Sinica, 2022, 49(6): 1181-1199.
[7]	ZHANG Kai, MA Mingying, WANG Ping, LI Yi, JIN Yan, SHENG Ling, DENG Ziniu, MA Xianfeng. Identification of HSP20 Family Genes in Citrus and Their Expression in Pathogen Infection Responses Citrus Canker [J]. Acta Horticulturae Sinica, 2022, 49(6): 1213-1232.
[8]	LIANG Chen, SUN Ruyi, XIANG Rui, SUN Yimeng, SHI Xiaoxin, DU Guoqiang, WANG Li. Genome-wide Identification of Grape GRF Family and Expression Analysis [J]. Acta Horticulturae Sinica, 2022, 49(5): 995-1007.
[9]	XIAO Xuechen, LIU Mengyu, JIANG Mengqi, CHEN Yan, XUE Xiaodong, ZHOU Chengzhe, WU Xingjian, WU Junnan, GUO Yinsheng, YEH Kaiwen, LAI Zhongxiong, LIN Yuling. Whole-genome Identification and Expression Analysis of SNAT,ASMT and COMT Families of Melatonin Synthesis Pathway in Dimocarpus longan [J]. Acta Horticulturae Sinica, 2022, 49(5): 1031-1046.
[10]	GAO Weilin, ZHANG Liman, XUE Chaoling, ZHANG Yao, LIU Mengjun, ZHAO Jin. Expression of E-type MADS-box Genes in Flower and Fruits and Protein Interaction Analysis in Chinese Jujube [J]. Acta Horticulturae Sinica, 2022, 49(4): 739-748.
[11]	LIU Mengyu, JIANG Mengqi, CHEN Yan, ZHANG Shuting, XUE Xiaodong, XIAO Xuechen, LAI Zhongxiong, LIN Yuling. Genome-wide Identification and Expression Analysis of GDSL Esterase/Lipase Genes in Dimocarpus longan [J]. Acta Horticulturae Sinica, 2022, 49(3): 597-612.
[12]	JIANG Cuicui, FANG Zhizhen, ZHOU Danrong, LIN Yanjuan, YE Xinfu. Identification and Expression Analysis of Sugar Transporter Family Genes in‘Furongli’（Prunus salicina） [J]. Acta Horticulturae Sinica, 2022, 49(2): 252-264.
[13]	WANG Zhiyu, CHANG Beibei, LIU Qi, CHENG Xiaofan, DU Xiaoyun, YU Xiaoli, SONG Laiqing, ZHAO Lingling. Study on Expression and Anthocyanin Accumulation of Solute Carrier Gene MdSLC35F2-like in Apple [J]. Acta Horticulturae Sinica, 2022, 49(11): 2293-2303.
[14]	HUANG Renwei, REN Yinghong, QI Weiliang, ZENG Rui, LIU Xinyu, DENG Binyan. Cloning of Mulberry MaERF105-Like Gene and Its Expression Under Drought Stress [J]. Acta Horticulturae Sinica, 2022, 49(11): 2439-2448.
[15]	BIAN Shicun, LU Yani, XU Wujun, CHEN Boqing, WANG Guanglong, XIONG Aisheng. Garlic Circadian Clock Genes AsRVE1 and AsRVE2 and Their Expression Analysis Under Osmotic Stress [J]. Acta Horticulturae Sinica, 2021, 48(9): 1706-1716.