Transcriptome and expression profile analysis of highly resistant and susceptible banana roots challenged with Fusarium oxysporum f. sp. cubense tropical race 4.

Banana wilt disease, caused by the fungal pathogen Fusarium oxysporum f. sp. cubense 4 (Foc4), is regarded as one of the most devastating diseases worldwide. Cavendish cultivar 'Yueyoukang 1' was shown to have significantly lower disease severity and incidence compared with susceptible cultivar 'Brazilian' in greenhouse and field trials. De novo sequencing technology was previously performed to investigate defense mechanism in middle resistant 'Nongke No 1' banana, but not in highly resistant cultivar 'Yueyoukang 1'. To gain more insights into the resistance mechanism in banana against Foc4, Illumina Solexa sequencing technology was utilized to perform transcriptome sequencing of 'Yueyoukang 1' and 'Brazilian' and characterize gene expression profile changes in the both two cultivars at days 0.5, 1, 3, 5 and 10 after infection with Foc4. The results showed that more massive transcriptional reprogramming occurs due to Foc4 treatment in 'Yueyoukang 1' than 'Brazilian', especially at the first three time points, which suggested that 'Yueyoukang 1' had much faster defense response against Foc4 infection than 'Brazilian'. Expression patterns of genes involved in 'Plant-pathogen interaction' and 'Plant hormone signal transduction' pathways were analyzed and compared between the two cultivars. Defense genes associated with CEBiP, BAK1, NB-LRR proteins, PR proteins, transcription factor and cell wall lignification were expressed stronger in 'Yueyoukang 1' than 'Brazilian', indicating that these genes play important roles in banana against Foc4 infection. However, genes related to hypersensitive reaction (HR) and senescence were up-regulated in 'Brazilian' but down-regulated in 'Yueyoukang 1', which suggested that HR and senescence may contribute to Foc4 infection. In addition, the resistance mechanism in highly resistant 'Yueyoukang 1' was found to differ from that in middle resistant 'Nongke No 1' banana. These results explain the resistance in the highly resistant cultivar and provide more insights in understanding the compatible and incompatible interactions between banana and Foc4.

RNA silencing suppressor Pns11 of rice gall dwarf virus induces virus-like symptoms in transgenic rice.

Transgenic rice (Oryza sativa) plants expressing the Pns11 protein of rice gall dwarf virus (RGDV) displayed multiple abnormal phenotypes, some of which were highly reminiscent of the symptoms observed in RGDV-infected rice. Further analysis indicated that the apparent alterations in plant growth and morphology were correlated with the expression levels of microRNA160, microRNA162, microRNA167, microRNA168, and the microRNA target OsARF8. Especially, the striking dwarfing phenotype depended on the high expression level of microRNA167. By analogy to other categories of plant viruses, the RNA silencing suppressors encoded by plant dsRNA viruses function as pathogenicity determinants. These findings significantly deepen our current mechanistic understanding of the RNA silencing suppressor (VSR) encoded by a dsRNA virus and provide additional evidence that interference with microRNA expression is a VSR function utilized by a diverse range of viruses.

[Main bacterial groups in banana soil under rotated and continuous cropping].

Banana wilt is the main disease in banana production, while banana-leek rotation can effectively control the occurrence of the disease. In order to understand the variations of soil bacterial groups under banana-leek rotation and banana continuous cropping, soil samples under these two cropping systems were collected to extract crude DNA, and the bacterial 16S rDNA in V3 region was amplified by PCR. The PCR products were then separated by DGGE, and the main different bands were sequenced and compared with the records of NCBI to identify the germs. Under banana-leek rotation, soil bacterial diversity was richer, and the main bacterial groups were Bacteroidetes, Proteobacteria, Actinobacteria, Chloroflexi, and Acidobacteria; while under banana continuous cropping, the soil bacterial diversity was somewhat decreased, and the main bacterial groups were Firmicutes, Proteobacteria, Actinobacteria, and Chloroflexi.

[Transgenic tobacco plants with ribosome inactivating protein gene cassin from Cassia occidentalis and their resistance to tobacco mosaic virus].

Cassin, the new gene of ribosome-inactivating protein (RIP) isolated from Cassia occidentalis, was inserted into expression vector pBI121 to produce plant expression vector pBI121-cassin (Figs.1, 2). pBI121-cassin was introduced into tobacco cultivar 'K326' by the Agrobacteriurm tumefaciens transformation method and more than 100 independent transformants were obtained. Southern blot hybridization analysis showed that a single gene locus was inserted into the chromosome of the transgenic tobacco lines (Fig.5) and PCR analysis of segregation population of progeny indicated that the inheritance of transgene was dominant in transgenic lines (Fig.4, Table 1). Results of RT-PCR and Northern blot hybridization analysis showed that transgene could be transcribed correctly (Figs.5, 6) . Three self-pollination lines of transgenic T(1) and T(2) were challenged with TMV at different concentration titers by mechanical inoculation. The transgenic lines exhibited different levels of resistance to TMV with the nontransgenic plants. After both titers of TMV concentration were inoculated, transgenic lines were considered as the highly resistant type with a delay of 4-13 d in development of symptoms and 10%-25% of test plants were infected, while nontransgenic control plants were susceptible typical symptoms on the newly emerged leaves (Table 2). One T(2) line, T(2)-8-2-1, was regarded as an immune type because it did not show any symptoms during 70 d and all plants were shown to be virus free by ELISA tests.