Molecular cloning and functional analysis of heat-tolerant nematode resistance genes from wild tomato (L. peruvianum LA2157)
Date Issued
2006
Date
2006
Author(s)
Ho, Hsiang-Ling
DOI
zh-TW
Abstract
Root-knot nematodes (Meloidogyne spp.) are important endoparasitic pests of numerous crop species. The feedings result in damages to tomato plants, including poor fruit yield, stunted growth, wilting and susceptibility to other pathogens, cause billions of dollars in agricultural losses each year. Tomato asscession Lycopersicon peruvianum LA2157 possesses heat-stable nematode resistance gene Mi-9, which is highly resistant to Mi-1-aviruent root-knot nematodes at 32 °C, was identified by Milligan et al.. However, it has not been cloned. In this experiment firstly, a 2-D gel analysis of protein patterns from infected roots was established timely. The most abundant protein spots were found at 24 hr after inoculation. Five differentially expressed proteins were picked up and identified by MALDI/MS, and two interesting proteins i.e. prosystemin and putative auxin response factor, were investigated further. Secondly, subtractive EST clones were produced by subtracting ESTs of control from those of 12-hr post-infecting roots. Total 384 ESTs were chosen for sequencing and analyzed. Among 361 valid ESTs, catalase isozyme2, heat stress transcription factors 30, transcription factor JERF1, cysteine proteinase inhibitor, thaumatin-like protein were further investigated. Northern blot analysis showed that all of these candidate genes were differentially expressed at 0, 4, 8, 12, and 24 hour after inoculation, suggesting that these genes work in different time points and in resistant pathway. Finally, virus-induced gene silencing method was used for functional analysis. The physiological functions in nematode-resistant molecular mechanism and relationship between Mi-9 of these candidate genes will be discussed.
Subjects
根瘤線蟲
番茄
抗線蟲基因
病毒誘導型基因靜默
root-knot nematode
tomato
nematode resistence gene
virus-induced gene silencing
Type
other
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