Agrobacterium-mediated production of transgenic rice plants expressing a chimeric α-amylase promoter/β-glucuronidase gene

We have successfully transferred and expressed a reporter gene driven by an -amylase promoter in a japonica type of rice (Oryza sativa L. cv. Tainung 62) using the Agrobacterium-mediated gene transfer system. Immature rice embryos (10–12 days after anthesis) were infected with an Agrobacterium strain carrying a plasmid containing chimeric genes of -glucuronidase (uidA) and neomycin phosphotransferase (nptII). Co-incubation of potato suspension culture (PSC) with the Agrobacterium inoculum significantly improved the transformation efficiency of rice. The uidA and nptII genes, which are under the control of promoters of a rice -amylase gene (Amy8) and Agrobacterium nopaline synthase gene (nos), respectively, were both expressed in G418-resistant calli and transgenic plants. Integration of foreign genes into the genomes of transgenic plants was confirmed by Southern blot analysis. Histochemical localization of GUS activity in one transgenic plant (R0) revealed that the rice -amylase promoter functions in all cell types of the mature leaves, stems, sheaths and roots, but not in the very young leaves. This transgenic plant grew more slowly and produced less seeds than the wild-type plant, but its R1 and R2 progenies grew normally and produced as much seeds as the wild-type plant. Inheritance of foreign genes to the progenies was also confirmed by Southern blot analysis. These data demonstrate successful gene transfer and sexual inheritance of the chimeric genes.     

Enigmatic double-stranded RNA in Japonica rice

We have found a linear, 16 kb, double-stranded RNA (dsRNA) in symptomless Japonica rice (Oryza sativa L.) that is not found in Indica rice (Oryza sativa L.). The dsRNA was detected in every tissue and at every developmental stage, and its copy number was approximately constant (about 20 copies/cell). Double-stranded RNA was also detected in two strains of Oryza rufipogon (an ancestor of O. sativa). Hybridization experiments indicated that the dsRNA of O. rufipogon was homologous but not identical to that of O. sativa. The sequence of about 13.2 kb of the dsRNA was determined and two open reading frames (ORFs) were found. The larger ORF (ORF B) was more than 12 351 nucleotides long and encoded more than 4 117 amino acid residues.     

再生稻褐飞虱为害损失测定和防治指标的研究

在福建,褐飞虱是再生稻的重要害虫,中进行了再生稻褐飞虱为害损失的测定,并对褐飞虱为害再生稻造成的产量损失结构进行了通径分析。根据虫量和产量损失的关系,结合现行稻谷价格,产量水平,防治费用等因素。确定允许为害损失率,同时建立了再生稻褐飞虱防治指标模型,按照防治指标模型,制定出不同产量水平的防治指标。     

Ar^＋,远红外激光,γ射线单一及复合处理水稻的诱变效果

用Ar~+、远红外两种激光和~(60)Co—r射线单一或复合处理两个籼稻品种的干种子,分析和比较了不同处理对水稻的当代生物学效应及处理二代的变异频率。结果表明,两种激光对当代的几个性状均表现为刺激效应,并有减轻,射线辐射损伤的作用,复合处理二代的变异频率和变异类型数明显高于相应的单一处理。说明复合处理是提高激光育种效果的有效方法。     

Factors controlling the population dynamics of the clonal helophyte Ranunculus lingua

Abstract. Two marginal and two central populations of the pseudo-annual aquatic plant Ranunculus lingua were studied over four years. The main purpose was to quantify potentially influential abiotic and biotic factors and to derive predictions about life-history differences between the populations. Variation in abundance and height of R. lingua ramets at different depths were related to water-level fluctuations, to abundance of other helophyes (emergent macrophytes), and to the occurrence of invertebrate grazing and fungal pathogens. Clear differences between marginal and central populations were shown in the depth distribution of ramet numbers and ramet heights, as well as in the dynamic patterns, where marginal populations had a higher flux of ramets. These patterns and regression analyses indicated that abiotic factors have a greater influence in marginal populations, whereas biotic factors are more important in central populations. It is suggested that marginal habitats for R. lingua would favour life-histories with a high reproductive capacity, whereas a large size of ramet would be the most important life-history feature in central habitats. This was supported by the fact that ramets in marginal populations, in spite of their smaller size, produced higher number of rhizomes than ramets in central populations. Variation in regional abundance was finally related to differences in demographic processes and dispersal potential between the populations.     

Calcineurin B-like interacting protein kinase 31 confers resistance to sheath blight via modulation of ROS homeostasis in rice

Sheath blight (ShB) severely threatens rice cultivation and production; however, the molecular mechanism of rice defence against ShB remains unclear. Screening of transposon Ds insertion mutants identified that Calcineurin B-like protein-interacting protein kinase 31 (CIPK31) mutants were more susceptible to ShB, while CIPK31 overexpressors (OX) were less susceptible. Sequence analysis indicated two haplotypes of CIPK31: Hap_1, with significantly higher CIPK31 expression, was less sensitive to ShB than the Hap_2 lines. Further analyses showed that the NAF domain of CIPK31 interacted with the EF-hand motif of respiratory burst oxidase homologue (RBOHA) to inhibit RBOHA-induced H₂O₂ production, and RBOHA RNAi plants were more susceptible to ShB. These data suggested that the CIPK31-mediated increase in resistance is not associated with RBOHA. Interestingly, the study also found that CIPK31 interacted with catalase C (CatC); cipk31 mutants accumulated less H₂O₂ while CIPK31 OX accumulated more H₂O₂ compared to the wild-type control. Further analysis showed the interaction of the catalase domain of CatC with the NAF domain of CIPK31 by which CIPK31 inhibits CatC activity to accumulate more H₂O₂.     

Upgrading the genome of an elite japonica rice variety Kongyu 131 for lodging resistance improvement

Developing a new rice variety requires tremendous efforts and years of input. To improve the defect traits of the excellent varieties becomes more cost and time efficient than breeding a completely new variety. Kongyu 131 is a high-performing japonica variety with early maturity, high yield, wide adaptability and cold resistance, but the poor-lodging resistance hinders the industrial production of Kongyu 131 in the Northeastern China. In this study, we attempted to improve the lodging resistance of Kongyu 131 from perspectives of both gene and trait. On the one hand, by QTL analysis and fine mapping we discovered the candidate gene loci. The following CRISPR/Cas9 and transgenic complementation study confirmed that Sd1 dominated the lodging resistance and favourable allele was mined for precise introduction and improvement. On the other hand, the Sd1 allelic variant was identified in Kongyu 131 by sequence alignment, then introduced another excellent allelic variation by backcrossing. Then, the two new resulting Kongyu 131 went through the field evaluation under different environments, planting densities and nitrogen fertilizer conditions. The results showed that the plant height of upgraded Kongyu 131 was 17%–26% lower than Kongyu 131 without penalty in yield. This study demonstrated a precise and targeted way to update the rice genome and upgrade the elite rice varieties by improving only a few gene defects from the perspective of breeding.