Guard cells of V. faba were prepared by sonicating epidermal Peels andtreated with ABA, mannitol and Ca2+.RNAs extracted from leaf tissue andguard cells were hybridized with RNAprobes of several representative RABgene...Guard cells of V. faba were prepared by sonicating epidermal Peels andtreated with ABA, mannitol and Ca2+.RNAs extracted from leaf tissue andguard cells were hybridized with RNAprobes of several representative RABgenes. An RNA with a sequence cognated to Pea dehydrin (psdhn1 ) was detected in water-stressed leaves and inthe guard cells treated with ABA for 1and 6 h (Figs. 1 & 2). No RNA cognate to osmotin gene was detected in either tissue preparation. No differencewas found between the transcripts ofRAB genes from whole leaves and thosefrom guard cells (Fig. 1). ABA, butnot Ca2+, was required for the formation of sequence cognated to psdhn1 inguard cells. Exogenous osmotun did notpromote the formation of the RNA sequence.展开更多
Rab proteins constitute the largest family of small GTPases,which play pivotal roles in intracellular membrane trafficking in all eukaryotes.A number of Rab genes have been identified in eukaryotes;however,very little...Rab proteins constitute the largest family of small GTPases,which play pivotal roles in intracellular membrane trafficking in all eukaryotes.A number of Rab genes have been identified in eukaryotes;however,very little information about these genes has been reported in insects.In the current study,for the first time we identified and characterized 27 Rab family genes from Locusta migratoria.Phylogenetic analysis and comparison of domain architecture indicated that Rab family genes are highly conserved among insect species.Tissue-dependent expression profiles indicated that expression of Rab genes was highest in the ovary,except for LmRab3,which was most highly expressed in hemolymph.The biological function of each Rab gene was investigated using RNA interference(RNAi).Double-stranded RNA targeting each Rab gene was injected into the hemocoel of nymphs and revealed that suppression of two Rab genes(LmRab5 and LmRab11A)caused 100%mortality.In addition,nymphs injected with dsLmRab5 exhibited severe phenotypic defects in the gastric caeca and midgut,while dsLmRab11A arrested the molting process.We then applied the RNAi of RNAi technique to test if silencing either of these two genes would affect the suppression of the lethal giant larvae(LmLgl)reporter gene and found that suppression of LmRab5 diminished the RNAi efficiency of LmLgl,whereas suppression of LmRab11A enhanced RNAi efficiency of LmLgl.These results indicate that Rab genes contribute differently to RNAi efficiency in different tissues.Our study provides a foundation for further functional investigations of Rab genes and their contributions to RNAi efficiency in L.migratoria.展开更多
文摘Guard cells of V. faba were prepared by sonicating epidermal Peels andtreated with ABA, mannitol and Ca2+.RNAs extracted from leaf tissue andguard cells were hybridized with RNAprobes of several representative RABgenes. An RNA with a sequence cognated to Pea dehydrin (psdhn1 ) was detected in water-stressed leaves and inthe guard cells treated with ABA for 1and 6 h (Figs. 1 & 2). No RNA cognate to osmotin gene was detected in either tissue preparation. No differencewas found between the transcripts ofRAB genes from whole leaves and thosefrom guard cells (Fig. 1). ABA, butnot Ca2+, was required for the formation of sequence cognated to psdhn1 inguard cells. Exogenous osmotun did notpromote the formation of the RNA sequence.
基金supported by National Natural Science Foundation of China(Grant No.31730074,31802018)2017 Special Talents Projects in Shanxi Province,China(201705D211027)Key Research and Development Program of Shanxi Province(201803D221004-5).
文摘Rab proteins constitute the largest family of small GTPases,which play pivotal roles in intracellular membrane trafficking in all eukaryotes.A number of Rab genes have been identified in eukaryotes;however,very little information about these genes has been reported in insects.In the current study,for the first time we identified and characterized 27 Rab family genes from Locusta migratoria.Phylogenetic analysis and comparison of domain architecture indicated that Rab family genes are highly conserved among insect species.Tissue-dependent expression profiles indicated that expression of Rab genes was highest in the ovary,except for LmRab3,which was most highly expressed in hemolymph.The biological function of each Rab gene was investigated using RNA interference(RNAi).Double-stranded RNA targeting each Rab gene was injected into the hemocoel of nymphs and revealed that suppression of two Rab genes(LmRab5 and LmRab11A)caused 100%mortality.In addition,nymphs injected with dsLmRab5 exhibited severe phenotypic defects in the gastric caeca and midgut,while dsLmRab11A arrested the molting process.We then applied the RNAi of RNAi technique to test if silencing either of these two genes would affect the suppression of the lethal giant larvae(LmLgl)reporter gene and found that suppression of LmRab5 diminished the RNAi efficiency of LmLgl,whereas suppression of LmRab11A enhanced RNAi efficiency of LmLgl.These results indicate that Rab genes contribute differently to RNAi efficiency in different tissues.Our study provides a foundation for further functional investigations of Rab genes and their contributions to RNAi efficiency in L.migratoria.
