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Digital Gene Expression Analysis of huanglongbing Affected Mandarins(citrus reticulata Blanco)In Response to Thermotherapy

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Horticultural Plant Journal 2021年第1期7卷 1-12页

作者： Ruixuan Pan Yanling Chen Xiaoling Deng Meirong Xu laboratory of citrus huanglongbing research Guangdong Key Laboratory of Microbial Signals and Disease ControlSouth China Agricultural UniversityGuangzhou 510642China

huanglongbing(HLB),which is mainly associated with“Candidatus Liberibacter asiaticus”(CLas),is currently threatening citrus production around the *** destructive century-old disease results in callose deposition in phloem plasmodesmata and starch overaccumulation in leaf *** treatment with high humidity has proven to be effective in eliminating the CLas titers in potted citrus *** study explored the curative mechanisms and impact of heat treatment(from 32 to 48°C with 75%±5%relative humidity,14h in light and 10h in dark for four days)on mandarin trees(citrus reticulata Blanco)that were severely infected by CLas using Solexa/Illumina’s digital gene expression(DGE)profiling.A total of 838 differentially expressed genes(DEGs)were identified,of which 510 were *** interaction analysis of the DEGs indicated that the thermotherapy of HLB trees downregulated the circadian clock related genes in chloroplasts and upregulated the RNA-editing process in the mitochondria,characterized by 141 genes encoding pentatricopeptide repeatcontaining proteins(PPR)being all ***,the expression of phenylpropanoid and flavonoid biosynthesis genes,and auxininduced genes were *** defense response pathway of the thermotherapy treated diseased trees was apparently disrupted,as indicated by the downregulation of LRR receptor-like serine/threonine-protein kinase FLS2 *** study improves our understanding of the curative mechanisms of heat treatment on HLB trees and CLas biological ***,this provides new insights on the involvement of RNA modification by PPR genes and changes in the circadian clock after the heat stress treatment of CLas-infected citrus.

关键词： citrus huanglongbing heat transcriptom differentially expressed genes

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The citrus huanglongbing Crisis and Potential Solutions

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Molecular Plant 2019年第5期12卷 607-609页

作者： Nian Wang citrus research and Education Center Department of Microbiology and Cell ScienceUniversity of Florida/lnstitute of Food and Agricultural SciencesLake AlfredFLUSA China-USA citrus huanglongbing Joint laboratory(A Joint laboratory of The University of Florida's Institute of Food and Agricultural Sciences and Gannan Normal University) National Navel Orange Engineering Research CenterGannan Normal UniversityGanzhouJiangxi 341000China

citrus is one of the top three fruit crops *** production is facing an unprecedented crisis due to huanglongbing(HLB,also known as citrus greening),which is the most devastating disease of *** was first reported in Asia a century *** is associated with Candidatus Liberibacter asiaticus(Las),***(Lam),and ***(Bove,2006),which belong to the gramegative *** them,Las is the most widely *** and Lam are vectored by Diaphorina citri Kuwayama(ACP),whereas Laf is transmitted by Trioza erytreae.

关键词： The citrus huanglongbing Crisis Potential Solutions citrus

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Curent Situation of “Candidatus Liberibacter asiaticus” in Guangdong,China,Where citrus huanglongbing Was First Described

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Journal of Integrative Agriculture 2012年第3期11卷 424-429页

作者： DENG Xiao-ling GAO Yi-di CHEN Jian-chi PU Xue-lian KONG Wei-wen LI Hua-ping laboratory of citrus huanglongbing research South China Agricultural University Guangzhou 510642 P.R. China San Joaquin Valley Agricultural Sciences Center USDA-ARS Parlier California 93648 USA School of Horticulture and Plant Protection Yangzhou University Yangzhou 225009 P.R. China

citrus huanglongbing （HLB,yellow shoot disease） was first observed in the coastal Chaoshan Plain of Guangdong Province,China,in the late 19th century based on descriptions of yellow shoot symptoms. “Candidatus Liberibacter asiaticus”has been considered as a putative pathogen associated with HLB since *** about the curent prevalence of this bacterium is important for HLB control in Guangdong and also provides useful reference for HLB study *** 2007,we collected HLB symptomatic citrus samples from 16 cultivars in 12 prefecture cities,mostly in the north and west regions of Guangdong,where major citrus fruits are currently *** the 359 samples collected,241 （67.1%） were positive for “***”,distributed in 15 out of the 16 cultivars from all 12 cities,indicating the widespread prevalence of “***” in Guangdong *** detection rates varied from 16.7 to 100% depending on location and *** detection rates were found in newer citrus cultivation cities among the previously less popular but now promoted *** reviewing the citrus management and pest control practice,we believe that infected nursery stocks play a key role in the current spread of “***”.

关键词： citrus huanglongbing Candidatus Liberibacter asiaticus

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An endolysin gene from Candidatus Liberibacter asiaticus confers dual resistance to huanglongbing and citrus canker

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Horticulture research 2023年第9期10卷 165-180页

作者： Lanzhen Xu Kaiqing Mo Danlu Ran Juanjuan Ma Lehuan Zhang Yijia Sun Qin Long Guojin Jiang Xiaochun Zhao Xiuping Zou Integrative Science Center of Germplasm Creation in Western China(CHONGQING)Science City Citrus Research InstituteSouthwest University/National Citrus Engineering Research CenterChongqing 400712China

The most damaging citrus diseases are huanglongbing(HLB)and citrus canker,which are caused by Candidatus Liberibacter asiaticus(CaLas)and Xanthomonas citri ***(Xcc),*** from bacteriophages are a possible option for disease resistance in plant ***,we report improvement of citrus resistance to HLB and citrus canker using the LasLYS1 and LasLYS2 endolysins from ***2 demonstrated bactericidal efficacy against several Rhizobiaceae bacteria and Xcc,according to inhibition zone *** two genes,driven by a strong promoter from Cauliflower mosaic virus,35S,were integrated into Carrizo citrange via Agrobacterium-mediated *** than 2 years of greenhouse testing indicated that LasLYS2 provided substantial and long-lasting resistance to HLB,allowing transgenic plants to retain low CaLas titers and no obvious symptoms while also clearing CaLas from infected plants in the long ***2 transgenic plants with improved HLB resistance also showed resistance to Xcc,indicating that LasLYS2 had dual resistance to HLB and citrus canker.A microbiome study of transgenic plants revealed that the endolysins repressed Xanthomonadaceae and Rhizobiaceae populations in roots while increasing Burkholderiaceae and Rhodanobacteraceae populations,which might boost the citrus defense response,according to transcriptome *** also found that Lyz domain 2 is the key bactericidal motif of LasLYS1 and *** endolysins with potential resistance to HLB and citrus canker were found based on the structures of LasLYS1 and ***,the work shed light on the mechanisms of resistance of CaLas-derived endolysins,providing insights for designing endolysins to develop broad-spectrum disease resistance in citrus.

关键词： citrus canker resistance

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Distribution pattern and titer of Candidatus Liberibacter asiaticus in periwinkle(Catharanthus roseus)

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Journal of Integrative Agriculture 2018年第11期17卷 2501-2508页

作者： LI Ya XU Mei-rong DAI Ze-han DENG Xiao-ling Agricultural College Guangdong Ocean University laboratory of citrus huanglongbing research/Guangdong Key laboratory of Microbial Signals and Disease Control South China Agricultural University

Candidatus Liberibacter asiaticus （CaLas）, an uncultured Gram-negative alphaproteobacterium, is the causal agent of huanglongbing （HLB） in citrus. CaLas resides in phloem sieve tubes and has been shown to be unequally distributed in different tissues. Although HLB is a disease of citrus plants, it has been demonstrated that periwinkle can serve as an experimental host of CaLas, which can be transmitted from citrus to periwinkle via the parasitic plant dodder （Cuscuta spp.）. To investigate the distribution of CaLas in various periwinkle tissues, the bacteria were transmitted from an infected periwinkle plant to healthy periwinkles by top-grafting. The movement of the inoculum and associated titer changes were observed over time in various tissues. CaLas could be detected in the leaves, main stems, and roots of infected periwinkle by conventional PCR, and in all three tissues a clear time-dependent change in CaLas titer was observed, with titer increasing soon after inoculation and then decreasing as disease symptoms became severe. The highest titer was found at 25, 35 and 35 days after inoculation in leaves, main stems and roots, respectively. The titer in leaves was much higher than in the main stems and roots at the same time point, and the spatial distribution of CaLas in the leaves, main stems and roots of infected periwinkle was uneven, similar to what has been shown in citrus. The results provide guidance for selecting the proper periwinkle tissues and sampling times for early detection of CaLas.

关键词： Candidatus Liberibacter asiaticus titer distribution Catharanthus roseus

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High genetic variation and recombination events in the vicinity of non-autonomous transposable elements from ‘Candidatus Liberibacter asiaticus'

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Journal of Integrative Agriculture 2015年第10期14卷 2002-2010页

作者： WANG Xue-feng CHEN Jiao-yue TAN Jin DUAN Suo DENG Xiao-ling CHEN Jian-chi ZHOU Chang-yong National citrus Engineering research Center Citrus Research Institute Chinese Academy of Agricultural Sciences Southwest University College of Plant Protection Southwest University citrus huanglongbing research Center South China Agricultural University San Joaquin Valley Agricultural Sciences Center (SJVASC) USDA-ARS

Two miniature inverted-repeat transposable elements（MITEs）, MCLas-A and MCLas-B, were recently identified from ‘Candidatus Liberibacter asiaticus＇ known to be associated with citrus huanglongbing（HLB, yellow shoot disease）. MCLas-A was suggested as an active MITE because of its mobility. The immediate upstream gene of the two MITEs was predicted to be a putative transposase. The goal of this study is to analyze the sequence variation in the upstream putative transposase of MITEs and explore the possible correlation between sequence variation of transposase gene and MITE activity. PCR and sequence analysis showed that 12 sequence types were found in six major amplicon types from 43 representative ‘Ca. L. asiaticus＇ isolates from China, the United States and Brazil. Out of the 12 sequence types, three（T4, T5-2, T6） were reported for the first time. Recombination events were found in the two unique sequence types（T5-2 and T6） which were detected in all Brazilian isolates. Notably, no sequence variation or recombination events were detected in the upstream putative transposase gene of MCLas-A, suggesting the conservation of the transposase gene might be closely related with the MITE activity. Phylogenetic analysis demonstrated two well supported clades including five subclades were identified, clearly reflecting the geographical origins of isolates, especially that of Ruili isolates, S？o Paulo isolates and a few Florida isolates.

关键词： huanglongbing ‘Candidatus Liberibacter asiaticus' miniature inverted-repeat transposable element

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Development of a sensitive and reliable droplet digital PCR assay for the detection of ‘Candidatus Liberibacter asiaticus'

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Journal of Integrative Agriculture 2018年第2期17卷 483-487页

作者： ZHONG Xi LIU Xue-lu LOU Bing-hai ZHOU Chang-yong WANG Xue-feng National citrus Engineering research Center Citrus Research Institute Chinese Academy of Agricultural Sciences/Southwest University Guangxi Key laboratory of citrus Biology Guangxi Academy of Specialty Crops

citrus huanglongbing（HLB, yellow shoot disease） is one of the most serious citrus diseases worldwide. To better improve the detection sensitivity, a droplet digital PCR（ddPCR） assay was developed for the rapid detection of ‘Candidatus Liberibacter asiaticus＇（Las）, the putative causal agent of HLB. The detection of sensitivity comparison using positive plasmid indicated that dd PCR was superior to quantitative PCR（qPCR） for detecting and quantifying Las at low concentrations. The Las detection of 40 field samples also showed that six of 13 asymptomatic samples（46.15%） with high Ct value（〉35） were positive by dd PCR. This methodology showed great potential for early HLB infection diagnosis.

关键词： citrus huanglongbing early diagnosis droplet digital PCR

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Mapping of QTLs and candidate genes associated with multiple phenotypic traits for huanglongbing tolerance in citrus

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Horticultural Plant Journal 2023年第4期9卷 705-719页

作者： Ming Huang Mikeal L. Roose Qibin Yu Ed Stover David G. Hall Zhanao Deng Frederick G.Gmitter Jr. Key laboratory of Plant Hormones and Development Regulation of Chongqing College of Life SciencesChongqing UniversityChongqing 401331China citrus research and Education Center University of FloridaLake AlfredFL 33850United States Department of Botany and Plant Sciences University of CaliforniaRiversideCA 92521United States United States Horticultural research laboratory Agricultural Research ServiceUnited States Department of AgricultureFort PierceFL 34945United States Gulf Coast research and Education Center University of FloridaWimaumaFL 33598United States

huanglongbing(HLB)is the most devastating disease for citrus *** Liberibacter asiaticus(C Las),vectored by Asian citrus psyllid(ACP,Diaphorina citri Kuwayama),is the most common pathogen causing the *** citrus varieties are highly susceptible to HLB,whereas trifoliate orange(Poncirus trifoliata)is considered highly tolerant to *** F1 segregating population and their parent trifoliate orange and sweet orange,which had been exposed to intense HLB pressure for three years,was evaluated for disease symptoms,ACP colonization,C Las titer and tree vigor repeatedly for two to three *** orange and sweet orange showed significant differences for most of the phenotypic traits,and the F1 population exhibited a large variation.A high-density SNP-based genetic map with 1402 markers was constructed for trifoliate orange,which exhibited high synteny and high coverage of its reference genome.A total of 26 quantitative trait locus(QTLs)were identified in four linkage groups LG-t6,LG-t7,LG-t8 and LG-t9,of which four QTL clusters exhibit a clear co-localization of QTLs associated with different *** genome-wide analysis of gene expression in response to C Las infection in‘Flying Dragon’and‘Larger-Flower DPI-50-7’trifoliate orange,85 differentially expressed genes were found located within the QTL *** them,seven genes were classified as defense or immunity protein which exhibited the highest transcriptional change after C Las *** results indicate a quantitative genetic nature of HLB tolerance and identified candidate genes that should be valuable for searching for genetic solutions to HLB through breeding or genetic engineering.

关键词： Candidatus liberibacter QTL mapping Phenotyping F1 population Poncirus citrus

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Diversity analysis of "Candidatus Liberibacter asiaticus" associated phage type in China

Diversity analysis of "Candidatus Liberibacter asiaticus" as...

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中国植物病理学会2015年学术年会

作者： ZHENG Zheng ZENG Yi-bing BAO Min-li XU Mei-rong DENG Xiao-ling citrus huanglongbing research laboratory South China Agricultural University

"Candidatus Liberibacter asiaticus"(CLas) is associated with citrus huanglongbing(HLB),one of the most damaging diseases in citrus production *** research has shown the presence of phage/prophage in CLas ***/prophages are important components of biological entities that directly contribute to the bacterial evolution and population *** prophage,FP1 and FP2,were been reported in CLas Floridian strain *** another Florida strain UF506,two prophages,SCI and SC connected in tandem,and their circular replication forms were ***1 and FP2 are basically identical to SCI and SC,respectively.A CLas Chinese strain gxpsy,which was from Guangxi province,was reported to have two prophages in tandem in the bacterial ***,the CLas Japanese strain Ishi-1 was reported to be none phage was ***,we sequenced a CLas strain(A4) from Guangdongprovince,where the citrus HLB was first described in *** analyses indicated that the A4 genome had a single prophage sharing similarity to *** better study the phage/prophage type in Chinese CLas population,eight PCR primer sets each specific to SC2 and SCI were *** preliminary experiment,60 CLas strains were collected from seven provinces,including Guangdong(24),Yunnan(12),Fujian(8),Hunan(6),Guangxi(2),Jiangxi(2),Zhejiang(2) and Hainan(4).Initial result indicated that SCI-type phage/prophage was only found in 2 provinces(Yunnan and Hainan) and SC2-type phage/prophage was found in 7 provinces(except Yunnan).Based on the specific primers PCR,the phage/prophage types of CLas population in China can be divided into four groups,single SCl-type,single SC2-type,none phage type and SC1-SC2 phage *** single SC1-type phage/prophages(SCl-type) were found in 15 strains(15/60,25%),including all 12 CLas Yunnan strains and 3 Hainan strains(3/4,75%).The single SC2-type phage/prophages(SC2-type)were found in 40 strains(40/60,66.7%),including 23 Guangdong strains(23/24,95.

关键词： Candidatus Liberibacter asiaticus specific phage/prophage type diversity

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Inhibition of trehalase affects the trehalose and chitin metabolism pathways in Diaphorina citri(Hemiptera:Psyllidae)

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Insect Science 2021年第3期28卷 718-734页

作者： Hai-Zhong Yu Yu-Ling Huang Zhan-Jun Lu Qin Zhang Hua-Nan Su Yi-Ming Du Long Yi Ba-Lian Zhong Ci-Xiang Chen College of Life Sciences Gannan Normal UniversityGanzhouJiangxiChina National Navel Orange Engineering research Center GanzhouJiangxiChina China-USA citrus huanglongbing Joint laboratory A Joint Laboratory of The University of Florida and Gannan Normal UniversityGanzhouJiangxiChina Fruit Bureau of Ganzhou GanzhouJiangxiChina

The Asian citrus psyllid,Diaphorina citri is the principal vector of huanglongbing,which transmits Candidatus Liberibacter *** is a key enzyme involved in trehalose hydrolysis and plays an important role in insect growth and *** specific functions of this enzyme in *** have not been *** this study,three trehalase genes(DcTreJ-1,DcTrel-2,and DcTre2)were identified based on the *** genome *** analysis showed that DcTrel-1 and DcTrel-2 are related to soluble trehalase,whereas DcTre2 is associated with membrane-bound *** expression analysis indicated that DcTrel-1 and DcTrel-2 had the highest expression levels in the head and wing,respectively,and DcTre2 had high expression levels in the fat ***,DcTrel-1 and DcTrel-2 expression levels were induced by 20-hydroxyecdysone and juvenile hormone III,but DcTre2 was *** expression levels of DcTrel-1,DcTrel-2,and DcTre2 were significantly up-regulated,which resulted in high mortality after treatment with *** activities and glucose contents were downregulated,but the trehalose content increased after treatment with *** addition,the expression levels of chitin metabolism-related genes significantly decreased at 24 and 48 h after treatment with ***,silencing of DcTrel-1,DcTrel-2,and DcTre2 reduced the expression levels of chitin metabolism-related genes and led to a malformed phenotype of *** results indicate that *** trehalase plays an essential role in regulating chitin metabolism and provides a new target for control of ***.

关键词： chitin metabolism Diaphorina citric RNA interference trehalase validamycin

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