dad-1, an endogenous programmed cell death suppressor in Caenorhabditis elegans and vertebrates.

Programmed cell death (apoptosis) is a normally occurring process used to eliminate unnecessary or potentially harmful cells in multicellular organisms. Recent studies demonstrate that the molecular control of this process is conserved phylogenetically in animals. The dad-1 gene, which encodes a novel 113 amino acid protein, was originally identified in a mutant hamster cell line (tsBN7) that undergoes apoptosis at restrictive temperature. We have identified a dad-1 homologue in Caenorhabditis elegans (Ce-dad-1) whose predicted product is > 60% identical to vertebrate DAD-1. A search of the sequence databases indicated that DAD-1-like proteins are also expressed in two plant species. Expression of either human dad-1 or Ce-dad-1 under control of a C.elegans heat-shock-inducible promoter resulted in a reduction in the number of programmed cell death corpses visible in C.elegans embryos. Extra surviving cells were present in these animals, indicating that both the human and C.elegans dad-1 genes can suppress developmentally programmed cell death. Ce-dad-1 was found to rescue mutant tsBN7 hamster cells from apoptotic death as efficiently as the vertebrate genes. These results suggest that dad-1, which is necessary for cell survival in a mammalian cell line, is sufficient to suppress some programmed cell death in C.elegans.     

dad-1, A Putative Programmed Cell Death Suppressor Gene in Rice

The human dad-1 cDNA homolog was isolated from rice plants.The amino acid sequence of the predicted protein product iswell conserved in both animals and plants. This rice dad-1 homologcan rescue the temperature-sensitive dad-1 mutants of hamstercells from apoptotic death, suggesting that the rice dad-1 homologalso functions as a suppressor for programmed cell death. (Received December 24, 1997; Accepted January 27, 1997)     

基于mtDNA COI基因序列的中国柑橘果实蝇的分子鉴定及柑橘大实蝇种群的遗传多样性分析

【目的】探明危害我国柑橘的实蝇种类以及柑橘大实蝇Bactrocera minax不同地理种群和不同寄主种群的遗传多样性。【方法】利用mtDNA COI基因对危害柑橘的果实蝇进行种类鉴定,采用MEGA软件对其中28个地理种群的535头果实蝇COI基因片段(约505 bp)序列进行比对,分析种间及种内遗传距离,构建系统发育树。使用DnaSP软件分析柑橘大实蝇不同地理种群和不同寄主种群的遗传多样性。【结果】从柑橘虫果内共鉴定出4种实蝇,分别为柑橘大实蝇B.minax、桔小实蝇B.dorsalis、蜜柑大实蝇B.tsuneonis和瑞丽果实蝇B.ruiliensis。这4种实蝇的种间遗传距离为0.0264～0.2410,种内遗传距离为0.0000～0.0140,种间与种内遗传距离没有重叠区域。单个柑橘虫果内一般仅有1种实蝇,极个别柑橘果实可同时被两种实蝇危害(4/43);在这些为害柑橘的实蝇种类中,以柑橘大实蝇的个体数量比例最大,占90.70%。柑橘大实蝇地理种群遗传多样性高,28个种群共有17个单倍型。【结论】柑橘大实蝇是所调查地区柑橘实蝇的绝对优势种,其种群遗传分化程度较高,扩散危害风险大。本研究结果对柑橘果实蝇类的监测和防控具有重要意义。     

Metabolomic analysis of citrus infection by 'candidatus liberibacter' reveals insight into pathogenicity

Huanglongbing (HLB), considered the most serious citrus disease in the world, is associated with the nonculturable bacterium 'Candidatus Liberibacter asiaticus' (Las). Infection of citrus by this pathogen leads to reduced plant vigor and productivity, ultimately resulting in death of the infected tree. It can take up to two years following initial infection before outward symptoms become apparent, making detection difficult. The existing knowledge gap in our understanding of Las and its pathogenesis leading to HLB has stymied development of treatments and methods to mitigate the pathogen's influence. To evaluate the influence of Las on fruit quality in both symptomatic and asymptomatic fruit, and gain further insight into the pathogenesis of the disease, a (1)H NMR metabolomics investigation, complemented with physicochemical and analyte-specific analyses, was undertaken. Comparison of the juice obtained from oranges gathered from Las(+) (symptomatic and asymptomatic) and Las(-) (healthy) trees revealed significant differences in the concentrations of sugars, amino and organic acids, limonin glucoside, and limonin. This study demonstrates differing metabolic profiles in the juice of oranges from Las(+) and Las(-) and proposes how Las may be able to evade citrus defense responses.     

Induction of a Citrus gene highly homologous to plant and yeast thi genes involved in thiamine biosynthesis during natural and ethylene-induced fruit maturation

Maturing citrus fruit undergo pigment changes which can be enhanced by exogenous ethylene. In order to identify genes induced by ethylene in citrus fruit peel, we cloned the gene c-thi1. mRNA corresponding to c-thi1 increased gradually in the peel during natural fruit maturation and in response to ethylene. GA₃ pretreatment reduced the inductive effect of ethylene. Levels of c-thi1 increased also in juice sacs but the effect of ethylene was much less prominent. c-thi1 is homologous to yeast and plant genes encoding for an enzyme belonging to the pathway of thiamine biosynthesis. The data suggest that thiamine is involved in citrus fruit maturation.     

Effects of Various Elicitors on the Transcription of a β-1,3-endoglucanase Gene in Citrus Fruit

Very little is yet known regarding the molecular mechanisms involved in pathogen defense responses in citrus fruit. Recently, a basic β-1,3-endoglucanase (EC 3.2.2.39) belonging to the pathogenesis-related (PR) group of proteins, has been purified from Citrus sinensis (L) Osbeck cv. `Valencia' orange callus. Specific antibodies raised against the purified protein were used to screen `Valencia' callus and flavedo cDNA expression libraries, and to isolate its corresponding cDNA, designated gns1. The gns1 gene encodes a predicted polypeptide of 336 amino acids with a molecular mass of 37.3 kDa and a basic pI of 9.19, and shares 55–65% identity with several other plant β-1,3-endoglucanase proteins. Hereby, we show that the expression of the gns1 gene is markedly induced by wounding and inoculation with Penicillium digitatum (Pers. Fr.) Sacc., and following treatments with various elicitors that induce fruit resistance against P. digitatum . These treatments include UV irradiation, application of jasmonic acid (JA), β-aminobutyric acid (BABA), Candida oleophila antagonist yeast cells and hot water rinsing and brushing. Overall, based on various RNA gel blot hybridizations, we assume that gns1 is most likely to be part of the molecular mechanisms involved in pathogen defense responses in citrus fruit. *     

Profiles of gene family members related to carotenoid accumulation in citrus genus

Citrus fruit are an important reservoir of carotenoids. Numerous studies have been carried out to identify and profile the members of gene families involved in carotenoid biosynthetic pathway to explain the diversity of coloration in citrus fruit. It was found that gene expression analysis could not always explain the changes in carotenoid content and composition, indicating that other unknown genes and mechanisms should be operative. This review summarizes and updates the current knowledge on gene families involved in the citrus carotenoid biosynthetic pathway and their roles on the regulation of carotenoid biosynthesis, as well as provides insightful questions leading to future experimentation.     

Nonhost status of Citrus sinensis cultivar valencia and C. paradisi cultivar ruby red to Mexican Anastrepha fraterculus (Diptera: Tephritidae)

Anastrepha fraterculus (Wiedemann) is recognized as a pest of citrus, apples, and blackberries in South America. In Mexico, it is mainly found in fruit of the family Myrtaceae and has never been reported infesting citrus. Here, we sought to determine whether females stemming from Mexican A. fraterculus populations (collected in the state of Veracruz) would lay eggs in 'Valencia' oranges and 'Ruby Red' grapefruit and, if so, whether larvae would hatch and develop. We worked under laboratory and seminatural conditions (i.e., gravid females released in fruit-bearing, bagged branches in a commercial citrus grove) and used Anastrepha ludens (Loew), a notorious pest of citrus, as a control species. Under laboratory conditions, A. ludens readily accepted both oranges and grapefruit as oviposition substrates, but A. fraterculus rarely oviposited in these fruit (but did so in guavas, a preferred host) and no larvae ever developed. Eggs were deposited in the toxic flavedo (A. fraterculus) and nontoxic albedo (A. ludens) regions. Field studies revealed that, as was the case in the laboratory, A. fraterculus rarely oviposited into oranges or grapefruit and that, when such was the case, either no larvae developed (oranges) or of the few (13) that developed and pupated (grapefruit), only two adults emerged that survived 1 and 3 d, respectively (5-17% of the time necessary to reach sexual maturity). In sharp contrast, grapefruit exposed to A. ludens yielded up to 937 pupae and adults survived for >6 mo. Therefore, the inability of Mexican A. fraterculus to successfully develop in citrus renders the status of Mexican A. fraterculus as a pest of citrus in Mexico as unsubstantiated.     

Straw Mulching Reduces the Harmful Effects of Extreme Hydrological and Temperature Conditions in Citrus Orchards

Extreme weather conditions with negative impacts can strongly affect agricultural production. In the Danjiangkou reservoir area, citrus yields were greatly influenced by cold weather conditions and drought stress in 2011. Soil straw mulching (SM) practices have a major effect on soil water and thermal regimes. A two-year field experiment was conducted to evaluate whether the SM practices can help achieve favorable citrus fruit yields. Results showed that the annual total runoff was significantly (P<0.05) reduced with SM as compared to the control (CK). Correspondingly, mean soil water storage in the top 100 cm of the soil profile was increased in the SM as compared to the CK treatment. However, this result was significant only in the dry season (Jan to Mar), and not in the wet season (Jul to Sep) for both years. Interestingly, the SM treatment did not significantly increase citrus fruit yield in 2010 but did so in 2011, when the citrus crop was completely destroyed (zero fruit yield) in the CK treatment plot due to extremely low temperatures during the citrus overwintering stage. The mulch probably acted as an insulator, resulting in smaller fluctuations in soil temperature in the SM than in the CK treatment. The results suggested that the small effects on soil water and temperature changes created by surface mulch had limited impact on citrus fruit yield in a normal year (e.g., in 2010). However, SM practices can positively impact citrus fruit yield in extreme weather conditions.     

An ecological interaction between citrus fruit, Penicillium moulds and Drosophila immigrans Sturtevant (Diptera: Drosophilidae)

Abstract. 1. Potential Drosophila breeding sites were collected from a fruit market, and the adults allowed to emerge from them.
2. D.immigrans and D.melanogaster were the species with the highest frequency of emergences from citrus fruit.
3. D.immigrans was especially associated with citrus fruits infected with Penicillium italicum or P.digitatum , two specialist moulds of citrus. D.melanogaster was more often found in uninfected fruit.
4. In the laboratory D.immigrans larvae survived better than D.melanogaster larvae on Penicillium-infected citrus fruit.
5. These adaptations suggest that D.immigrans may have originally evolved as a citrus specialist, becoming a domestic species as these fruits were exploited commercially.