Isolation and characterization of differentially expressed genes in the mycelium and fruit body of Tuber borchii
Isolation and characterization of differentially expressed genes in the mycelium and fruit body of Tuber borchii
复制标题
DOI：
10.1128/aem.68.9.4574-4582.2002
复制
发表时间：
2002-09-01
影响因子：
4.4
通讯作者：
Martin, F
中科院分区：
文献类型：
--
作者：
Lacourt, I;Duplessis, S;Martin, F
块菌从营养菌丝体到子实体的转变需要分化过程，这导致由不同细胞和组织类型组成的可食用子实体（子囊）。在这些发育过程中差异表达的基因的鉴定可以大大有助于更好地了解块菌形态建成。利用白色块菌营养菌丝体RNA构建了一个cDNA文库，并对214个cDNA进行了测序。高达58%的表达序列标签对应于已知基因。大多数鉴定的序列代表管家蛋白，即，参与基因或蛋白质表达、细胞壁形成、初级和次级代谢以及信号传导途径的蛋白质。利用营养菌丝体和子囊菌的cDNA探针筛选了171个cDNA阵列，以鉴定子实体调控基因。营养菌丝体和子实体轴承15或70%的成熟孢子的信号比较显示高达33%的研究基因的表达水平的显着差异。通过RNA印迹分析证实了六个高度调节基因的表达水平。谷氨酰胺合成酶、5-氨基乙酰丙酸合成酶、异柠檬酸裂解酶、硫氧还蛋白、葡聚糖1，3-β-葡萄糖苷酶和UDP-葡萄糖：甾醇葡萄糖基转移酶的表达高度上调，表明氨基酸生物合成、乙醛酸循环途径和细胞壁合成在形态发生过程中发生显著改变。
The transition from vegetative mycelium to fruit body in truffles requires differentiation processes which lead to edible fruit bodies (ascomata) consisting of different cell and tissue types. The identification of genes differentially expressed during these developmental processes can contribute greatly to a better understanding of truffle morphogenesis. A cDNA library was constructed from vegetative mycelium RNAs of the white truffle Tuber borchii, and 214 cDNAs were sequenced. Up to 58% of the expressed sequence tags corresponded to known genes. The majority of the identified sequences represented housekeeping proteins, i.e., proteins involved in gene or protein expression, cell wall formation, primary and secondary metabolism, and signaling pathways. We screened 171 arrayed cDNAs by using cDNA probes constructed from mRNAs of vegetative mycelium and ascomata to identify fruit body-regulated genes. Comparisons of signals from vegetative mycelium and fruit bodies bearing 15 or 70% mature spores revealed significant differences in the expression levels for up to 33% of the investigated genes. The expression levels for six highly regulated genes were confirmed by RNA blot analyses. The expression of glutamine synthetase, 5-aminolevulinic acid synthetase, isocitrate lyase, thioredoxin, glucan 1,3-beta-glucosidase, and UDP-glucose:sterol glucosyl transferase was highly up-regulated, suggesting that amino acid biosynthesis, the glyoxylate cycle pathway, and cell wall synthesis are strikingly altered during morphogenesis.