甜橙SWEET2a促进柑橘溃疡病菌侵染

doi:10.16420/j.issn.0513-353x.2021-0323

摘要/Abstract

摘要：

柑橘溃疡病菌（Xanthomonas citri subsp. citri,Xcc）侵染感病种质冰糖橙叶片后,在侵染部位形成典型的火山口凸起症状,但在抗病种质枸橼C-05叶片侵染部位呈现逐渐褐色坏死,未形成溃疡病典型症状。基于前期冰糖橙和枸橼C-05叶片接种Xcc的转录组测序结果,分析鉴定的20个甜橙SWEET受Xcc诱导表达变化,结果显示SWEET2a和SWEET17d在冰糖橙中受Xcc诱导上调表达,SWEET12b在枸橼C-05中受Xcc诱导上调表达;实时荧光定量PCR验证,SWEET2a在冰糖橙叶片中受Xcc诱导高表达而在枸橼C-05叶片中表达变化不显著;SWEET2a在不同柑橘种质同源基因编码的氨基酸序列相似性为94.6%,其启动子顺式作用元件在冰糖橙和枸橼C-05中的种类和数量存在较大差异。冰糖橙和枸橼C-05叶片瞬时过表达结果显示,SWEET2a促进Xcc的繁殖,且拟南芥过表达转基因株系接种毒性菌Pst.DC3000（Pseudomonas syringae tomato DC3000）后,单位叶面积菌含量显著高于野生型。SWEET2a蛋白部分定位于质膜,表明SWEET2a可能被Xcc劫持用于跨膜运输糖类物质至胞外,供给Xcc在细胞间隙定殖所需要的能量。

关键词: 柑橘, 柑橘溃疡病菌, Xcc, SWEET2a, 基因功能

Abstract:

After the infection of citrus canker pathogen Xanthomonas citri subsp. citri（Xcc）,infected the leaves of the susceptible germplasm‘Bingtang’Sweet Orange,it formed typical crater bulge symptoms at the infected site,but gradually showed brown necrosis at the infected site of the leaves of the resistant germplasm Citron C-05,without typical symptoms of canker disease. Based on the transcriptome sequencing analysis of‘Bingtang’Sweet Orange and Citron C-05. leaves inoculated with Xcc,the expression of 20 CsSWEET induced by Xcc was compared. The results showed that SWEET2a and SWEET17d were up-regulated by Xcc in‘Bingtang’Sweet Orange,and SWEET12b was up-regulated by Xcc in Citron C-05;qRT-PCR verification only SWEET2a induced by Xcc high expression in ‘Bingtang’Sweet Orange and no significant change in Citron C-05,The amino acids sequence similarity of SWEET2a in different citrus germplasm homologs was 94.6%. The cis-acting elements of the promoter in‘Bingtang’Sweet Orange and Citron C-05 were significantly different in types and quantities. The transient overexpression of SWEET2a in‘Bingtang’Sweet Orange and Citron C-05 showed that SWEET2a promoted the reproduction of Xcc,and the transgenic Arabidopsis thaliana lines inoculated with the virulence strain Pst.DC3000（Pseudomonas syringae tomato DC3000）was significantly higher than that of the wild type. The SWEET2a protein is partially localized to the plasma membrane,suggesting that SWEET2a may be hijacked by Xcc for transmembrane transport of sugar extracellular to supply Xcc with energy needed for intercellular colonization.

Key words: citrus, Xcc, Citron C-05, SWEET2a, gene function

中图分类号:

S666

麻明英, 郝晨星, 张凯, 肖桂华, 苏翰英, 文康, 邓子牛, 马先锋. 甜橙SWEET2a促进柑橘溃疡病菌侵染[J]. 园艺学报, 2022, 49(6): 1247-1260.

MA Mingying, HAO Chenxing, ZHANG Kai, XIAO Guihua, SU Hanying, WEN Kang, DENG Ziniu, MA Xianfeng. CsSWEET2a Promotes the Infection of Xanthomonas citri subsp. citri[J]. Acta Horticulturae Sinica, 2022, 49(6): 1247-1260.

图/表 11

表1 引物列表

Table 1 The List of Primers

基因 Gene	引物名称 Primer ID	引物序列（5′-3′） Primer sequence	退火温度/℃ T_m	产物大小/ bp Size
SWEET17d	orange1.1t04897-qPCR-F1	AATTTCATTTTTGGGCTTCTAGGCA	58	105
SWEET17d	orange1.1t04897-qPCR-R1	AAATTCCTCCGTTGATCTACGCTGT
SWEET12b	orange1.1t02627-qPCR-F1	CACGTCTTCGTCTTCTCGGA	58	89
SWEET12b	orange1.1t02627-qPCR-R1	GCGTACGACCAGCCTCATAA
SWEET2a	Cs2g04140-qPCR-F1	TCGCTACAGTGAATTCAGTTGGAGC	60	174
SWEET2a	Cs2g04140-qPCR-R1	GCCGCCCATTGGAGTCAA
EF1a		GTAACCAAGTCTGCTGCCAAG
EF1a		GACCCAAACACCCAACACATT
SWEET2a	Cs2g04140-KpnⅠ-F	GGGGTACCATGTCTTCAGTTGGGATTTC	60	1 771
SWEET2a	Cs2g04140- KpnⅠ-R	CCGGTACCTGCAAATGAATCGATCAG
	Cs2g04140-F	TCTGGAACTCAAGCATTCCGACTTT	55	581
	p1300-eYFP-R	CCGGACACGCTGAACTTGT

图1 冰糖橙和枸橼C-05叶片注射接种Xcc和Xoo的症状

Fig. 1 Symptoms of Xcc and Xoo inoculation in‘Bingtang’Sweet Orange and Citron C-05

图2 冰糖橙和枸橼C-05的SWEET家族基因受Xcc诱导差异表达分析

Fig. 2 Differential expression analysis of SWEET family in ‘Bingtang’ Sweet Orange and Citron C-05 induced by Xcc

图3 冰糖橙和枸橼C-05 SWEETs受Xcc和Xoo诱导的表达量变化双尾t检验（* P < 0.05）。.

Fig. 3 Relative expression of SWEETs induced by Xcc and Xoo in‘Bingtang’Sweet Orange and Citron C-05 Two-tailed student’s t-test（* P < 0.05）.

图4 SWEET2a编码的氨基酸序列差异比对 BTC：冰糖橙;JY C-05：枸橼C-05;CGXY：长果香橼;Cg：柚;SJG：金柑;PTJY：普通枸橼。

Fig. 4 Sequence difference alignment of amino acids encoded by SWEET2a BTC：‘Bingtang’ Sweet Orange;JY C-05：Citron C-5;CGXY：Changguo Citron;Cg：Citrus Grandis;SJG：Fortunella Hindsii;PTJY：Common Citron.

图5 不同枸橼种质SWEET2a编码的氨基酸序列差异比对 JY C-05：枸橼C-05;AGXY：矮果香橼;PTJY：普通枸橼;YXY：园香橼;CGXY：长果香橼;DNXY：丹娜香橼;MGJY：美国枸橼;NCXY：南川香橼;YSXY：野生香橼;XXY：小香橼。

Fig. 5 Sequence difference alignment of amino acids encoded by SWEET2a in different Citron medica JY C-05：Citron C-5;AGXY：Aiguo Citron;PTJY：Common Citron;YXY：Round Citron;CGXY：Changguo Citron;DNXY：Danna Citron;MGJY：American Citron;NCXY：Nanchuan Citron;YSXY：Wild Citron;XXY：Small Citron.

表2 SWEET2a启动子顺式作用元件分析

Table 2 The Cis-acting regulatory elements in promoter of SWEET2a

顺式元件 cis-Element	功能预测 Function prediction	序列 Sequence	起始位/bp Start position
顺式元件 cis-Element	功能预测 Function prediction	序列 Sequence	冰糖橙 ‘Bingtang’Sweet Orange	枸橼C-05 Citron C-05
ARE	厌氧诱导Anaerobic induction	AAACCA	+ 212	-905
			+ 1 046	-933
			+ 1 489
TCA-element	水杨酸响应Salicylic acid responsiveness	TCAGAAGAGG	+ 1 924
MBS	MYB结合位点参与干旱诱导	CAACTG	-1 626	+ 181
	MYB binding site involved in drought-inducibility			-611
				-1 422
LTR	低温诱导Low-temperature responsiveness	CCGAAA	-649	-623
CAT-box	分生组织表达Meristem expression	GCCACT	+ 198	+ 1 571
			-1 155
			+ 1 768
TC-rich repeats	防御和应激反应Defense and stress responsiveness	GTTTTCTTAC	+ 1 478
ABRE	脱落酸响应Abscisic acid responsiveness	ACGTG	+ 185
			+ 186
			-681
			+ 855
			-1 122
			-1 124
			+ 1 125
			-1 221
CCAAT-box	MYBHv1结合位点MYBHv1 binding site	CAACGG	-1 500	-1 067
O2-site	Zein新陈代谢调节Zein metabolism regulation	GATGATGTGG		-356
O2-site		GATGATGTGG		+ 399
GCN4_motif	胚乳表达Endosperm expression	TGAGTCA		+ 44
TGACG-motif	茉莉酸甲酯响应MeJA-responsiveness	TGACG		-1 419
TATC-box	赤霉素响应Gibberellin responsiveness	TATCCCA		-341

图6 SWEET2a在本氏烟叶片的表达特征 A：注射含有35S::SWEET2a:eYFP的本氏烟;B：注射含有35S::SWEET2a:eYFP与细胞质膜定位标签基因PM-RB的本氏烟叶片质壁分离。黄色箭头指示SWEET2a荧光表达,蓝色箭头指示细胞壁,白色箭头指示PM-RB荧光,红色箭头指示为二者融合荧光。

Fig. 6 The expression patten of SWEET2a in Nicotiana benthamiana A：The Nicotiana benthamiana leaf containing 35S::SWEET2a:eYFP was injected;B：After injection of 35S::SWEET2a:eYFP and cytoplasmic membrane localization gene PM-RB,the plasma wall of Nicotiana benthamiana leaf was isolated. The yellow arrow indicates the fluorescence expression of SWEET2a;The blue arrow indicates the location of the cell wall;The white arrow indicates the fluorescence expression of PM-RB;The red arrows indicate merge expression of the two.

图7 瞬时过表达SWEET2a冰糖橙和枸橼C-05叶片接种Xcc后15 d症状（A）和0 ~ 4 d Xcc生长量（B） A：SWEET2a瞬时过表达的冰糖橙叶片接种后15 d症状;B：瞬时过表达后,冰糖橙中Xcc生长量。双尾t检验（* P < 0.05）。

Fig. 7 The symptom of Xcc in leaves of‘Bingtang’Sweet Orange and Citron C-05 after transient overexpression of SWEET2a at 15 d（A）and the growth of Xcc at 0-4 d（B） A：The symptoms of transient overexpression of SWEET2a in‘Bingtang’Sweet Orange leaves in 15 d;B：The growth of Xcc in‘Bingtang’ Sweet Orange after transient overexpression. Two-tailed Student’s t-test（* P < 0.05）.

图8 拟南芥T1代转基因植株OE-SWEET2a验证

Fig. 8 Verification of T1 generation transgenic plants OE-SWEET2a in Arabidopsis thaliana

图9 OE-SWEET2a 接种Pst.DC3000后3 d症状（A）和病菌含量（B） A：接种Pst. DC3000后3 d症状;B：单位叶面积Pst. DC3000含量;Wild Type：野生型拟南芥对照。双尾t检验（* p < 0.05）。

Fig. 9 Symptoms（A）and pathogen content（B）of OE-SWEET2a at 3 days after inoculation with Pst.DC3000 A：Symptoms of Pst. DC3000 in 3 d;B：Pst. DC3000 content per unit leaf area;WT：Wild Type. Two-tailed Student’s t-test（* P < 0.05）.

参考文献 39

[1]	Behlau F, Belasque J J, Bergamin F A, Graham J H, Leite J R P, Gottwald T R. 2008. Copper sprays and windbreaks for control of citrus canker on young orange trees in southern Brazil. Crop Protection, 27 (3-5):807-813. doi: 10.1016/j.cropro.2007.11.008 URL
[2]	Rhonda C M, Steinfath M, Lisec J, Becher M, Witucka-Wall H, TorjéK O, Fiehn O, Eckardt A, Willmitzer L, Selbig J, Altmann T. 2007. The metabolic signature related to high plant growth rate in Arabidopsis thaliana. Proceedings of the National Academy, 104 (11):4759-4764.
[3]	Chandran D. 2015. Co-option of developmentally regulated plant SWEET transporters for pathogen nutrition and abiotic stress tolerance. Iubmb Life, 67 (7):461-471. doi: 10.1002/iub.1394 pmid: 26179993
[4]	Chardon F, Bedu M, Calenge F, Klemens P A W, Spinner L, Clement G, Chietera G, Léran S, Ferrand M, Lacombe B, Loudet O, Dinant S, Bellini C, Neuhaus H E, Daniel-Vedele F, Krapp A. 2013. Leaf fructose content is controlled by the vacuolar transporter SWEET17. Current Biology, 23 (8):697-702. doi: 10.1016/j.cub.2013.03.021 pmid: 23583552
[5]	Chen H Y, Huh J H, Yu Y C, Ho L H, Chen L Q, Tholl D, Frommer W B, Guo W J. 2015. The Arabidopsis vacuolar sugar transporter SWEET 2 limits carbon sequestration from roots and restricts pythium infection. The Plant Journal, 83 (6):1046-1058. doi: 10.1111/tpj.12948 URL
[6]	Chen L Q, Hou B H, Lalonde S, Takanaga H, Hartung M L, Qu X Q, Guo W, Kim J, Underwood W, Chaudhuri B, Chermak D, Antony G, White F F, Somerville S C, Mudgett M B, Frommer W B. 2010. Sugar transporters for intercellular exchange and nutrition of pathogens. Nature, 468 (7323):527-532. doi: 10.1038/nature09606 URL
[7]	Chen L Q, Qu X Q, Hou B H, Sosso D, Osorio W S, Fernie A R, Frommer W B. 2012. Sucrose efflux mediated by SWEET proteins as a key step for phloem transport. Science, 335:207-211. doi: 10.1126/science.1213351 URL
[8]	Chen L Q. 2014. SWEET sugar transporters for phloem transport and pathogen nutrition. New Phytologist, 201 (4):1150-1155. doi: 10.1111/nph.12445 URL
[9]	Chong J L, Piron M C, Meyer S, Merdinoglu D, Bertsch C, Mestre P. 2014. The SWEET family of sugar transporters in grapevine:VvSWEET4 is involved in the interaction with Botrytis cinerea. Journal of Experimental Botany, 65:6589-6601. doi: 10.1093/jxb/eru375 URL
[10]	Chu Z H, Fu B Y, Yang H, Xu C G, Li Z K, Sanchez A, Park Y J, Bennetzen J L, Zhang Q F, Wang S P. 2006. Targeting xa13,a recessive gene for bacterial blight resistance in rice. Theoretical and Applied Genetics, 112:455-461. doi: 10.1007/s00122-005-0145-6 URL
[11]	Cohn Megan, Morbitzer Robert, Bart Rebecca S, Hou B H, Shybut M, Frommer W B, Dahlbeck D, Lahaye T, Gomez M, Staskawicz B J. 2014. Xanthomonas axonopodis Virulence is promoted by a transcription activator-like effector-mediated induction of a SWEET sugar transporter in Cassava. Molecular Plant-Microbe Interactions, 27 (11):1186-1198. doi: 10.1094/MPMI-06-14-0161-R pmid: 25083909
[12]	Cox K L, Meng F H, Wilkins K E, Li F J, Wang P, Booher N J, Carpenter C D, Chen L Q, Zheng H, Gao X Q, Zheng Y, Fei Z J, Yu J Z, Isakeit T, Wheeler T, Frommer W B, He P, Bogdanove A J, Shan L B. 2017. TAL effector driven induction of a SWEET gene confers susceptibility to bacterial blight of cotton. Nature Communications, 8:15588. doi: 10.1038/ncomms15588 URL
[13]	Das A K. 2003. Citrus canker-a review. J Appl Hort, 5 (1):52-60.
[14]	Ference C M, Baldwin E A, Manthey J A, Jones J B. 2020. Inhibitory extracts of calamondin leaves associated with precipitous decline of Xanthomonas citri subsp. citri populations. European Journal of Plant Pathology, 156 (2):451-461. doi: 10.1007/s10658-019-01894-w URL
[15]	Frank B R, Kristen A L, David M B. 2012. SWEET as sugar:new sucrose effluxers in plants. Molecular Plant, 5 (4):766-768. doi: 10.1093/mp/sss054 pmid: 22815540
[16]	Gao Y, Wang Z, Kumar V, Xu X F, Yuan D P, Zhu X F, Li T Y, Ji B L, Xuan Y H. 2018. Genome-wide identification of the SWEET gene family in wheat. Gene, 642:284-292. doi: 10.1016/j.gene.2017.11.044 URL
[17]	Geng Yanqiu, Dong Xiaochang, Zhang Chunmei. 2021. Recent progress of sugar transporter in horticultural crops. Acta Horticulturae Sinica, 48 (4):676-688. (in Chinese)
	耿艳秋, 董肖昌, 张春梅. 2021. 园艺作物糖转运蛋白研究进展. 园艺学报, 48 (4):676-688.
[18]	Gottwald T R, Graham J H, Civerolo E L, Barrett H C, Hearn C J. 1993. Differential host range reaction of citrus and citrus relatives to citrus canker and citrus bacterial spot determinated by leaf mesophyll susceptiblity. Plant Disease, 77 (10):1004-1009. doi: 10.1094/PD-77-1004 URL
[19]	Guo W J, Nagy R, Chen H Y, Pfrunder S, Yu Y C, Santelia D, Frommer W B, Martinoia E. 2014. SWEET17,a facilitative transporter,mediates fructose transport across the tonoplast of Arabidopsis roots and leaves. Plant Physiology, 164 (2):777-789. doi: 10.1104/pp.113.232751 URL
[20]	Hu Y, Zhang J L, Jia H G, Sosso D, Li T, Frommer W B, Yang B, White F F, Wang N, Jones J B. 2014. Lateral organ boundaries 1 is a disease susceptibility gene for citrus bacterial canker disease. Proc Natl Acad Sci USA, 111(4):E521-529.
[21]	Jean C R T, Stroock A D. 2017. Phloem loading through plasmodesmata a biophysical analysis. Plant Physiology, 175:904-915. doi: 10.1104/pp.16.01041 URL
[22]	Joan D, Emily G, Christina K, Francoise S P, Casieri L, Wipf D. 2012. Sugar transporters in plants and in their interactions with fungi. Trends in Plant Science, 17 (7):413-422. doi: 10.1016/j.tplants.2012.03.009 URL
[23]	Lalonde S, Wipf D, Frommer W B. 2004. Transport mechanisms for organic forms of carbon and nitrogen between source and sink. Annu Rev Plant Biol, 55:341-372. pmid: 15377224
[24]	Li Qiang, Qi Jingjing, Dou Wanfu, Qin Xiujuan, He Yongrui, Chen Shanchun. 2020. Overexpression of CsNBS-LRR in citrus confers bacterial canker resistance by regulating SA signaling pathway. Acta Horticulturae Sinica, 47 (5):817-826. (in Chinese)
	李强, 祁静静, 窦万福, 秦秀娟, 何永睿, 陈善春. 2020. 柑橘超量表达CsNBS-LRR 通过SA 信号途径增强对溃疡病抗性. 园艺学报, 47 (5):817-826.
[25]	Liu Q S, Meng Y, Zhou Y, Li X H, Xiao J H, Wang S P. 2011. A paralog of the MtN3/saliva family recessively confersrace-speciﬁc resistance to Xanthomonas oryzae in rice. Plantcell Environ, 34:1958-1969.
[26]	Liu X Z, Zhang Y, Yang C, Tian Z H, Li J X. 2016. AtSWEET4,a hexose facilitator,mediates sugar transport to axial sinks and affects plant development. Scientific Reports, 6:1-12. doi: 10.1038/s41598-016-0001-8 URL
[27]	Ma Zhimin, Duan Yu, Xu Jianjian, Bin Yu, Zhou Changyong, Song Zhen. 2021. The rapid detection of Xanthomonas citri ssp. citri(Xcc)based on recombinase polymerase amplification(RPA)assay. Acta Horticulturae Sinica, 48 (3):590-599. (in Chinese)
	马志敏, 段玉, 许建建, 宾羽, 周常勇, 宋震. 2021. 基于重组酶聚合酶扩增技术(RPA)的柑橘溃疡病菌检测方法. 园艺学报, 48 (3):590-599.
[28]	Qi Jingjing, Dou Wanfu, Zhang Qingwen, Hu Anhua, Chen Shanchun, Lei Tiangang, Peng Aihong, Xu Lanzhen, Yao Lixiao, He Yongrui, Li Qiang. 2020. Interacting protein screening and analysis of CsAP2-09—a citrus bacterial canker related transcription factor. Acta Horticulturae Sinica, 47 (3):432-444. (in Chinese)
	祁静静, 窦万福, 张庆雯, 胡安华, 陈善春, 雷天刚, 彭爱红, 许兰珍, 姚利晓, 何永睿, 李强. 2020 柑橘抗溃疡病转录因子CsAP2-09互作蛋白筛选与分析. 园艺学报, 47 (3):432-444.
[29]	Qi Jingjing, Qin Xiujuan, Xie Yu, Chen Shanchun, He Yongrui, Li Qiang. 2021. Correlation analysis of citrus catalase gene CsKat01and citrus canker disease. Acta Horticulturae Sinica, 48 (1):26-36. (in Chinese)
	祁静静, 秦秀娟, 谢宇, 陈善春, 何永睿, 李强. 2021. 过氧化氢酶基因CsKat01与柑橘溃疡病相关性分析. 园艺学报, 48 (1):26-36.
[30]	Schaad N W, Postnikova E, Lacy G, Sechler A, Agarkova I, Stromberg P E, Stromberg V K, Vidaver A K. 2006. Emended classification of xanthomonad pathogens on citrus. Syst Appl Microbiol, 29 (8):690-695. doi: 10.1016/j.syapm.2006.08.001 URL
[31]	Schubert T S, Rizvi S A, Sun X A, Gottwald T R, Graham J H, Dixon W N. 2001. Meeting the challenge of eradicating citrus canker in Florida-Again. Plant Disease, 85 (4):340-356.
[32]	Slewinski T L. 2011. Diverse functional roles of monosaccharide transporters and their homologs in vascular plants:a physiological perspective. Molecular Plant, 4 (4):641-662. doi: 10.1093/mp/ssr051 pmid: 21746702
[33]	Smeekens S, Ma J K, Hanson J, Rolland F. 2010. Sugar signals and molecular networks controlling plant growth. Curr Opin Plant Biol, 13 (3):274-279. doi: 10.1016/j.pbi.2009.12.002 pmid: 20056477
[34]	Streubel J, Pesce C, Hutin M, Koebnik R, Boch J, Szurek B. 2013. Five phylogenetically close rice SWEET genes confer TAL effector-mediated susceptibility to Xanthomonas oryzae pv. oryzae. New Phytologist, 200 (3):808-819. doi: 10.1111/nph.12411 pmid: 23879865
[35]	Swarup S, Yinong Y, Kingsley M T, Gabriel D W. 1992. An Xanthomonas citri pathogenicity gene,pthA,pleiotropically encodes gratuitous avirulence on Nonhosts. Molecular Plant-Microbe Interactions, 5 (3):204-213. pmid: 1421509
[36]	Tang D J, He Y Q, Feng J X, He B R, Jiang B L, Lu G T, Chen B S, Tang J L. 2005. Xanthomonas campestris pv. campestris possesses a single gluconeogenic pathway that is required for virulence. Journal of bacteriology, 187 (17):6231-6237. doi: 10.1128/JB.187.17.6231-6237.2005 URL
[37]	Yang B, Sugio A, White F F. 2006. Os8N3 is a host disease-susceptibility gene for bacterial blight of rice. Proc Natl Acad Sci, 103 (27):10503-10508. doi: 10.1073/pnas.0604088103 URL
[38]	Zhou J H, Peng Z, Long J Y, Sosso D, Liu B, Eom J S, Huang S, Liu S Z, Cruz C V, Frommer W B, White F F, Yang B. 2015. Gene targeting by the TAL effector pthXo2 reveals cryptic resistance gene for bacterial blight of rice. Plant Journal, 82:632-643. doi: 10.1111/tpj.12838 URL
[39]	Zhu Zhi-mei, Tan Li-mei, Xu Jing, Fu Hong-yan, Hu Zhe, Gong Lei, Yang Gui-bing, Wang Ping, Ma Xian-feng, Deng Zi-niu. 2021. Evaluation of citron genotypes for the resistance to citrus canker. China Fruits,(1):43-49. (in Chinese)
	朱志媚, 谭李梅, 徐静, 符红艳, 胡哲, 龚蕾, 杨贵兵, 王萍, 马先锋, 邓子牛. 2021. 枸橼种质对柑橘溃疡病的抗性鉴定. 中国果树,(1):43-49.