Departamento de Ciencia de los Alimentos, Instituto de Agroquímica y Tecnología de Alimentos-CSIC, Apartado de Correos 73, Burjassot, E46100-Valencia, Spain.
BMC Plant Biol. 2010 Aug 31;10:194. doi: 10.1186/1471-2229-10-194.
Postharvest losses of citrus fruit due to green mold decay, caused by the fungus Penicillium digitaum, have a considerable economic impact. However, little is known about the molecular processes underlying the response of citrus fruit to P. digitatum.
Here we describe the construction of a subtracted cDNA library enriched in citrus genes preferentially expressed in response to pathogen infection followed by cDNA macroarray hybridization to investigate gene expression during the early stages of colonization of the fruit's peel by P. digitatum. Sequence annotation of clones from the subtracted cDNA library revealed that induction of secondary and amino acid metabolisms constitutes the major response of citrus fruits to P. digitatum infection. Macroarray hybridization analysis was conducted with RNA from either control, wounded, ethylene treated or P. digitatum infected fruit. Results indicate an extensive overlap in the response triggered by the three treatments, but also demonstrated specific patterns of gene expression in response to each stimulus. Collectively our data indicate a significant presence of isoprenoid, alkaloid and phenylpropanoid biosynthetic genes in the transcriptomic response of citrus fruits to P. digitatum infection. About half of the genes that are up-regulated in response to pathogen infection are also induced by ethylene, but many examples of ethylene-independent gene regulation were also found. Two notable examples of this regulation pattern are the genes showing homology to a caffeine synthase and a berberine bridge enzyme, two proteins involved in alkaloid biosynthesis, which are among the most induced genes upon P. digitatum infection but are not responsive to ethylene.
This study provided the first global picture of the gene expression changes in citrus fruit in response to P. digitatum infection, emphasizing differences and commonalities with those triggered by wounding or exogenous ethylene treatment. Interpretation of the differentially expressed genes revealed that metabolism is redirected to the synthesis of isoprenes, alkaloids and phenylpropanoids.
由青霉(Penicillium digitaum)引起的绿霉腐烂会导致柑橘果实采后损失,造成相当大的经济影响。然而,人们对柑橘果实对青霉的反应的分子过程知之甚少。
在这里,我们描述了一个富含柑橘基因的消减 cDNA 文库的构建,这些基因在受到病原体感染后优先表达,然后通过 cDNA 宏阵列杂交来研究青霉浸染柑橘果皮早期阶段的基因表达。从消减 cDNA 文库中克隆的序列注释表明,次生代谢和氨基酸代谢的诱导构成了柑橘果实对青霉感染的主要反应。用来自对照、受伤、乙烯处理或青霉感染的果实的 RNA 进行宏阵列杂交分析。结果表明,三种处理触发的反应有很大的重叠,但也表明了每种刺激的特定基因表达模式。总的来说,我们的数据表明,在柑橘果实对青霉感染的转录组反应中存在大量的异戊二烯、生物碱和苯丙烷生物合成基因。约一半因病原体感染而上调的基因也被乙烯诱导,但也发现了许多与乙烯无关的基因调控的例子。这种调控模式的两个显著例子是与咖啡因合酶和小檗碱桥酶同源的基因,这两种蛋白都参与生物碱的生物合成,是青霉感染后诱导最多的基因之一,但对乙烯没有反应。
这项研究提供了柑橘果实对青霉感染反应的基因表达变化的第一张全景图,强调了与受伤或外源乙烯处理触发的变化的差异和共同点。差异表达基因的解释表明,代谢被重新定向合成异戊二烯、生物碱和苯丙烷。