Interaction between endophytic bacteria from citrus plants and the phytopathogenic bacteria Xylella fastidiosa, causal agent of citrus-variegated chlorosis

AIMS: To isolate endophytic bacteria and Xylella fastidiosa and also to evaluate whether the bacterial endophyte community contributes to citrus-variegated chlorosis (CVC) status in sweet orange (Citrus sinensis [L.] Osbeck cv. Pera). METHODS AND RESULTS: The presence of Xylella fastidiosa and the population diversity of culturable endophytic bacteria in the leaves and branches of healthy, CVC-asymptomatic and CVC-symptomatic sweet orange plants and in tangerine (Citrus reticulata cv. Blanco) plants were assessed, and the in vitro interaction between endophytic bacteria and X. fastidiosa was investigated. There were significant differences in endophyte incidence between leaves and branches, and among healthy, CVC-asymptomatic and CVC-symptomatic plants. Bacteria identified as belonging to the genus Methylobacterium were isolated only from branches, mainly from those sampled from healthy and diseased plants, from which were also isolated X. fastidiosa. CONCLUSIONS: The in vitro interaction experiments indicated that the growth of X. fastidiosa was stimulated by endophytic Methylobacterium extorquens and inhibited by endophytic Curtobacterium flaccumfaciens. SIGNIFICANCE AND IMPACT OF THE STUDY: This work provides the first evidence of an interaction between citrus endophytic bacteria and X. fastidiosa and suggests a promising approach that can be used to better understand CVC disease.     

Citrus replant problem in Iraq II. Possible role of allelopathy

The role of allelopathy in citrus replant problems was investigated in Iraq. The failure of citrus seedlings to grow normally in old citrus orchards was not caused by differences between old and non-citrus soils in electrical conductivity, pH, organic matter, soil texture and those minerals tested. Extracts of soil collected from old citrus orchards significantly reduced the growth of sour orange seedlings. Extracts and decaying sour orange roots reduced the growth of sour orange seedlings as did extracts of non-senescent sour orange leaves and decaying senescent leaves. Thus it appears that allelopathy is at least partly involved in the citrus replant problem.     

Citrus orchard evapotranspiration: Comparison between eddy covariance measurements and the FAO-56 approach estimates

Abstract

The aim of this study was to use the FAO-56 single and dual crop coefficient approaches to estimate actual evapotranspiration (ET_a) over an irrigated citrus orchard under drip and flood irrigations in Marrakech, Morocco. The results showed that, by using crop coefficients suggested in the FAO-56 paper, the performance of both approaches was poor for two irrigation treatments. The Root Mean Squared Error (RMSE) between measured and simulated ET_a values over the citrus orchard under drip irrigation was about 1.43 and 1.27 mm/day for the single and dual approaches, respectively, while the corresponding statistics for the orchard irrigated by the flooding technique was 1.87 and 2.48 mm/day. After determination of the appropriate values of the crop coefficient (K_c) based on eddy covariance measurements of ET_a, the performance of both approaches greatly improved. The obtained K_c values were lower than the FAO-56 values by about 20%. The low K_c values obtained reflect the practice of drip irrigation for one field and the low value of cover fraction for the other field. Additionally, the efficiency of the irrigation practices was investigated by comparing the measured K_c for two fields. The results showed that a considerable amount of water was lost by direct soil evaporation from the citrus orchard irrigated by flooding technique.     

Genome sequencing and CRISPR/Cas9 gene editing of an early flowering Mini‐Citrus (Fortunella hindsii)

Hongkong kumquat (Fortunella hindsii) is a wild citrus species characterized by dwarf plant height and early flowering. Here, we identified the monoembryonic F. hindsii (designated as ‘Mini‐Citrus’) for the first time and constructed its selfing lines. This germplasm constitutes an ideal model for the genetic and functional genomics studies of citrus, which have been severely hindered by the long juvenility and inherent apomixes of citrus. F. hindsii showed a very short juvenile period (~8 months) and stable monoembryonic phenotype under cultivation. We report the first de novo assembled 373.6 Mb genome sequences (Contig‐N50 2.2 Mb and Scaffold‐N50 5.2 Mb) for F. hindsii. In total, 32 257 protein‐coding genes were annotated, 96.9% of which had homologues in other eight Citrinae species. The phylogenomic analysis revealed a close relationship of F. hindsii with cultivated citrus varieties, especially with mandarin. Furthermore, the CRISPR/Cas9 system was demonstrated to be an efficient strategy to generate target mutagenesis on F. hindsii. The modifications of target genes in the CRISPR‐modified F. hindsii were predominantly 1‐bp insertions or small deletions. This genetic transformation system based on F. hindsii could shorten the whole process from explant to T₁ mutant to about 15 months. Overall, due to its short juvenility, monoembryony, close genetic background to cultivated citrus and applicability of CRISPR, F. hindsii shows unprecedented potentials to be used as a model species for citrus research.     

DNA barcoding of genus Toxoptera Koch (Hemiptera: Aphididae): Identification and molecular phylogeny inferred from mitochondrial COI sequences

Abstract Identification of aphid species is always difficult due to the shortage of easily distinguishable morphological characters. Aphid genus Toxoptera consists of species with similar morphology and similar to Aphis in most morphological characters except the stridulatory apparatus. DNA barcodes with 1 145 bp sequences of partial mitochondrial cytochrome‐coxidase I (COI) genes were used for accurate identification of Toxoptera. Results indicated mean intraspecific sequence divergences were 1.33%, whereas mean interspecific divergences were greater at 8.29% (0.13% and 7.79% if T. aurantii 3 and T. aurantii 4 are cryptic species). Sixteen samples were distinguished to four species correctly by COI barcodes, which implied that DNA barcoding was successful in discrimination of aphid species with similar morphology. Phylogenetic relationships among species of this genus were tested based on this portion of COI sequences. Four species of Toxoptera assembled a clade with low support in maximum‐parsimony (MP) analysis, maximum‐likelihood (ML) analysis and Bayesian phylogenetic trees, the genus Toxoptera was not monophyletic, and there were two sister groups, such as T. citricidus and T. victoriae, and two clades of T. aurantii which probably presented cryptic species in the genus.     

Reprogramming of Stem Cell Activity to Convert Thorns into Branches

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中国优质水果资源的分布与适宜生态环境

根据农业部在80年代两次组织评选出的全国189个优质水果产地的生态环境资料,用微型电子计算机系统建立数据库,统计分析柑桔、苹果和梨优质产品的构成比例、产区分布地域及其适宜的环境指标和主栽品种的生态适应性,为果树良种区域化栽培与选育提供依据。     

巴氏新小绥螨对柑桔全爪螨处理的枳橙叶片挥发物的行为反应

【目的】研究巴氏新小绥螨Neoseiulus barkeri对柑桔全爪螨Panonychus citri及刺吸式昆虫为害柑桔叶片释放的挥发物的行为反应,揭示巴氏新小绥螨的嗅觉反应特点。【方法】采用GC-MS顶空进样法对枳橙叶片常见挥发性化合物、针刺枳橙叶片及其柑桔全爪螨雌成螨取食枳橙叶片挥发物进行定性分析,确定每类化合物的相对保留指数,构建枳橙叶片常见及受害挥发性化合物特征谱库。利用嗅觉测定技术分析巴氏新小绥螨对枳橙叶片挥发物的行为反应。【结果】针刺处理和柑桔全爪螨取食影响枳橙叶片挥发物的组成和含量。两种方法处理时枳橙叶片释放的主要物质为α-蒎烯、水芹烯、4-异丙基甲苯。随着处理加重,增量释放的物质为:cis-π-罗勒烯、月桂烯、柠檬烯、异松油烯,减量表达的物质为2-乙基-1-乙醇和十一烷。在10~(-2)、10~(-4)、10~(-6)和10~(-8)g/m L浓度下,正己醛、正壬醛、乙酸辛酯和正庚醛对巴氏新小绥螨有强烈的引诱作用(P>0.05);月桂烯在10~(-2)、10~(-4)和10~(-6)g/m L浓度下对巴氏新小绥螨有强烈的引诱作用(P>0.05);正壬醇和正辛醇随着浓度增加,对巴氏新小绥螨的引诱作用降低;苯甲醛对巴氏新小绥螨的引诱作用较弱。【结论】巴氏新小绥螨对柑桔全爪螨及刺吸式口器昆虫为害柑桔叶片释放出的挥发物各组分具有不同的行为反应,柑桔及其刺吸式害虫生境中的嗅觉线索在巴氏新小绥螨的寄主定位和生境选择中起着重要作用。