Proteomic maps of subcellular protein fractions of the Asian citrus psyllid Diaphorina citri,the vector of citrus huanglongbing

The Asian citrus psyllid Diaphorina citri Kuwayama (Hemiptera: Liviidae) is an insect vector that transmits the bacterial pathogen Candidatus Liberibacter asiaticus (CLas) associated with the destructive citrus disease, citrus huanglongbing (HLB). Currently, D. citri is the major target in HLB management, although insecticidal control and disruption of the D. citri–CLas interactions both face numerous challenges. The present study reports the subcellular proteomic profiles of D. citri, encompassing the three main subcellular protein fractions: cytosol, mitochondria and microsomes. After optimization, subcellular proteins of both high and low abundance are obtained by two‐dimensional gel electrophoresis (2‐DE). A total of 1170 spots are detected in the 2‐DE gels of the three subcellular fractions. One hundred and sixty‐four differentially expressed proteins are successfully identified using liquid chromatography‐dual mass spectroscopy. An efficient protocol for subcellular protein fractionation from D. citri is established and a clear protein separation is achieved with the chosen protein fractionation protocol. The identified cytosolic proteins are mainly metabolic enzymes, whereas a large portion of the identified proteins in the mitochondrial and microsomal fractions are involved in ATP biosynthesis and protein metabolism, respectively. Protein–protein interaction networks are predicted for some identified proteins known to be implicated in pathogen–vector interactions, such as actin, tubulin and ATP synthase, as well as insecticide resistance, such as the cytochrome P450 superfamily. The findings should provide useful information to help identify the mechanism responsible for the CLas–D. citri interactions and eventually contribute to D. citri control.     

SPE-HPLC-DAD法同时检测柑橘药用资源中黄烷酮类和川陈皮素成分

为检测柑橘类5种药用资源中芸香柚皮苷、柚皮苷、橙皮苷、柚皮素和橙皮素等5个黄烷酮物质和川陈皮素的含量。实验用70%乙醇水溶液分别浸提化橘红、陈皮、青皮、橘络和橘核,提取液经稀释后利用C18固相萃取柱除杂和浓缩,再以0.5%醋酸水溶液和甲醇为流动相进行反相梯度洗脱,用串联二极管阵列检测器(DAD)扫描紫外光谱做定性分析,并分别在283、285、290、335nm波长处做定量检测。在该条件下6个类黄酮成分均实现基线分离;外标法定量,线性相关性好(R2≥0.9998);加标回收率为95.83%~103.56%,相对标准偏差为2.90%~8.78%。含量分析结果显示:芸香柚皮苷10.49mg/g、柚皮素0.327mg/g、橙皮素0.129mg/g在青皮中含量最高,橙皮苷10.78mg/g、川陈皮素1.74mg/g在陈皮中含量最高,柚皮苷19.20mg/g在化橘红中含量最高。该方法适用于柑橘药用资源中微量类黄酮成分的准确定性和定量检测。     

植物激素在柑桔育苗中的应用

本文概述了在柑桔种苗生产中,植物激素对砧木种子萌发、嫁接成活、砧木苗和嫁接苗生长以及扦插生根的影响。     

Early events in Agrobacterium-mediated genetic transformation of citrus explants

BACKGROUND AND AIMS: Genetic transformation of plants relies on two independent but concurrent processes: integration of foreign DNA into plant cells and regeneration of whole plants from these transformed cells. Cell competence for regeneration and for transformation does not always fall into the same cell type/developmental stage, and this is one of the main causes of the so-called recalcitrance for transformation of certain plant species. In this study, a detailed examination of the first steps of morphogenesis from citrus explants after co-cultivation with Agrobacterium tumefaciens was performed, and an investigation into which cells and tissues are competent for regeneration and transformation was carried out. Moreover, the role of phytohormones in the co-cultivation medium as possible enhancers of gene transfer was also studied. METHODS: A highly responsive citrus genotype and well-established culture conditions were used to perform a histological analysis of morphogenesis and cell competence for transformation after co-cultivation of citrus epicotyl segments with A. tumefaciens. In addition, the role of phytohormones as transformation enhancers was investigated by flow cytometry. KEY RESULTS: It is demonstrated that cells competent for transformation are located in the newly formed callus growing from the cambial ring. Conditions conducive to further development of this callus, such as treatment of explants in a medium rich in auxins, resulted in a more pronounced formation of cambial callus and a slower shoot regeneration process, both in Agrobacterium-inoculated and non-inoculated explants. Furthermore, co- cultivation in a medium rich in auxins caused a significant increase in the rate of actively dividing cells in S-phase, the stage in which cells are more prone to integrate foreign DNA. CONCLUSIONS: Use of proper co-cultivation medium and conditions led to a higher number of stably transformed cells and to an increase in the final number of regenerated transgenic plants.     

Interactions between citrus psylla,Trioza erytreae (Hem. Triozidae), and spiders in an unsprayed citrus orchard in the transvaal lowveld

During a two year survey, a total of 3,054 spiders represented by 21 families were sampled in an unsprayed citrus orchard in the Transvaal Lowveld. Numerically the Salticidae was the dominant family (34.4%) followed by the Theridiidae (21.9%), Thomisidae (11.9%), Araneidae (7.9%), Clubionidae (7.0%) and the Tetragnathidae (3.7%). Eighteen species of spiders were observed to prey on citrus psylla,Trioza erytreae (Del Guercio) (Hemiptera: Triozidae), while six species trap nymphs and adults under their retreats and webs. There were significant positive correlations between the weekly psylla populations and the weekly populations of web-building spiders and wandering spiders present one to four weeks later but no significant correlation between the weekly spider populations and the weekly psylla populations present one to five weeks later. This seems to indicate that while spiders are unable to keep citrus psylla populations at acceptable low levels, they may contribute in reducing their numbers.      

桔园内多种天敌对桔全爪螨的控制作用

本文应用二次回归旋转组合实验设计统计方法,对桔园内主要天敌食螨瓢虫(Stethorus che-ngi)、长须螨(Agistemus exsertus citri)、尼氏钝绥螨(Amblyseius nichols)和普通钝绥螨(Amb-lyseius vulgaris)对桔全爪螨(Panongchus citri)的控制作用,以及天敌与天敌之间、天敌与害螨之间的相互作用进行了研究。得出了这4种天敌对桔全爪螨的捕食量模型。并对模型进行主效分析,表明桔全爪螨自身密度的大小对天敌的总捕食量影响最大。在这4种天敌中对总的捕食量影响最大的是食螨瓢虫,其次为长须螨,尼氏钝绥螨再次之,普通钝绥螨的影响最小。对各天敌间交互作用的分析表明,天敌个体大、活动能力强、捕食量大,他们的种内种间干扰程度也大,特别是食螨瓢虫种内种间干扰十分明显。其次为长须螨,而尼氏钝绥螨和普通钝绥螨的种内种间干扰较弱,在密度较低时,他们还表现出相互促进的作用。最后本文还讨论了这4种天敌能有效地控制桔全爪螨的最大密度,以及在该密度下各种天敌密度的最佳组合。     

Immature embryo rescue,culture and seedling development of acid citrus fruit derived from interploid hybridization

Interploid sexual hybridizations were completed in 2001 and 2002 between seven lemon (Citrus limon(L.) Burm. f.) varieties, Key lime (C. aurantifolia (Cristm.) Swing), Palestine sweet lime (C. imettioides Tan.), Lakeland limequat (C. aurantifolia x Fortunella japonica (Thumb.) Swing.), and Etrog citron (C. medica L.) as diploid progenitors and four allotetraploid somatic hybrids (Key lime + Valencia orange, Hamlin orange + Femminello lemon, Valencia orange + Rough lemon, and Valencia orange+ Femminello lemon) in efforts to generate improved seedless triploid acid fruit hybrids. Efficient recovery of triploid progeny from such crosses requires embryo rescue to avoid embryo abortion due to endosperm failure. Germination of rescued genetically diverse immature embryos was induced on two culture media (EME and Gamborgs B5), with two sucrose concentrations (50 or 70 g l^–1). All media contained 0.5 g l^–1 malt extract and 4.50 M GA₃. Germination of globular, heart and torpedo shaped embryos (defined as small embryos) was significantly (p < 0.05) affected by medium and genotype. Gamborgs medium induced 82.89% germination. Of germinated embryos, 11–65% developed into normal plants with differences among crosses. Cotyledonary embryos (defined as immature embryos with fully developed cotyledons) germinated and developed into normal plants at higher rates than less-developed embryos. In efforts to improve the efficiency of plant recovery, small embryos from Todo el año × HF and Lisbon × HF crosses conducted during 2002 were rescued and cultured on three media (MS, Gamborgs, and RMA) for comparison. Media did not significantly affect the proportion of normal plant recovery.     

Distribution of Phytophthora citrophthora R.E. Sm. and E.H. Sm. and P. parasitica Dastur within citrus orchards in Kerman province,Iran

In this research, distribution of Phytophthora species were determined in Kerman Province (Bam, Shahdad and Arzuiyeh) during 2004–2007. The Phytophthora species were isolated from infected root, crown and soil. Root and crown pieces were washed and cultured on a CMA-PARPH medium. The isolation from infected soil was performed by bating method using citrus leaves. It was identified by morphological and several physiological characteristics. Distribution of species was determined by recording the number of isolates recovered from samples from each city. In this study, from 220 soil samples collected from 52 citrus orchards, 80 isolates of Phytophthora were recovered. Among of all isolates of Phytophthora, P. parasitica and P. citrophthora were the most important species of causal agent of gummosis on Citrus sp. Distribution of P. citrophthora was highest in Arzuiyeh and lowest in Bam and Shahdad cities, while distribution of P. parasitica was highest in Bam and Shahdad cities. The majority of soil samples contained only P. parasitica, but a few of percentage samples containing P. citrophthora alone and both of fungi in cites samples.     

Morphology,life cycle and management of two invasive subspecies of Papilio demoleus (Lepidoptera: Papilionidae): A review

Papilio demoleus L., also known as the citrus butterfly or chequered swallowtail, is a common species in the Asia-Pacific region, with two of its subspecies (P. d. demoleus and P. d. malayanus) posing significant threats to the citrus industry in Asia and other regions of the world. Its predominant natural host plants are the Rutaceae for the invasive northern subspecies and Fabaceae for the southern subspecies. The morphology and development of this species are reasonably well studied, although further information is required on its natural mortality rates and environmental tolerance in order to accurately predict its future potential spread throughout the Middle East and Central Americas. Synthetic pyrethroids such as deltamethrin are highly effective for the control of P. demoleus larvae. However, better control is likely to be found using an integrated approach to pest management, incorporating techniques such as biocontrol agents, microbial pesticides and phytopesticides, in conjunction with synthetic pesticides where required.     

The biology and phenology of the citrus whitefly, Dialeurodes citri, on citrus in the Coastal Plain of Israel

The citrus whitefly, Dialeurodes citri (Ashmead) (Homoptera: Aleyrodidae) (CWF), is a polyphagous citrus pest, originating in S. E. Asia, which was first discovered in the Western Galilee region of Israel in 1975. The larval instars of CWF secrete honeydew, and the sooty mould fungus, which develops on the honeydew, causes damage to the tree and its fruit. In observations made in Israel between 1987 and 1991 CWF usually completed three generations per year, although a small proportion of the population had two or four generations. From October to April a combination of shortened photoperiod and reduced temperature arrest the development of CWF in the fourth larval instar. The average length of the life cycle from egg to adult emergence was 65 days in the first generation (April–June), 52 days in the second generation (July–August) and 8 months in the third generation (September–April). Under laboratory conditions of L16: D8 and 25 °C, the mean developmental time was 54 days, and the duration of the egg, first larval instar, second larval instar, third larval instar, fourth larval instar and putative pupa (red eyes) were 12.1, 6.5, 5.5, 7.6, 18, and 4.7 days, respectively. Adults prefer to lay their eggs on young, fully developed citrus leaves, but also oviposit on older leaves. The inner region and shaded sides of the tree are preferred for oviposition.