Identification of Growth-promoting Bacteria from Rhizosphere of Pastures and Their Effects on Growth of Lotus corniculatus L.

[Objectives] This study was conducted to explore the interaction between nitrogen-fixing and phosphate-solubilizing strains and the optimal combination of different functional strains,in order to provide a theoretical basis for the development of PGPR compound fertilizers suitable for local environment.[Methods] In this study,16S rDNA gene sequence analysis was used to identify fast-growing and competitive strains from pasture nodules and rhizosphere soils in Guizhou Province,and three representative Rhizobia and phosphorus-solubilizing bacteria were chosen for the test of bacterial combination when reducing 50% of nitrogen and 30% of phosphorus.The effects of different strain combinations on the plant height,root length,aboveground and underground biomass of Lotus corniculatus L.were investigated,and the total nitrogen and total phosphorus contents of the plants were determined.[Results] The mixed bacterial agents could promote the increase of root biomass,and the effects of A1,A3,B3 and C3 were the most obvious.The fresh weight and dry weight of the roots of L.corniculatus increased by 30.35%-168.45% and 26.43%-180.00%,respectively,and A3,B3,B2 and C3 had the best effects.The total phosphorus content of the plants increased by 12.79%-55.25% compared with the CK2;and most of the bacterial agents with significant growth-promoting effects showed decreased total nitrogen contents,while those with non-significant growth-promoting effects showed significantly-increased total nitrogen contents,which were not as much as the CK1.Comprehensively,the most productive combination was C3,namely R27-2 Rhinohizobium fredii and P33-3 Stenotrophomonas rhizophila.[Conclusions] This study can provide a theoretical basis for the production and promotion of bacterial fertilizers.

Construction of RNAi Expression Vector Targeting Vernalizational Gene FaCONSTANS in Tall Fescue

[ Objective ] This study aimed to construct RNAi expression vector targeting vemalizational gene FaCONSTANS （GenBank accession number： GU214996） in tall rescue. [ Method] A 145 bp long Arabidopsis actin gene intron was inserted into the expression vector to construct an intermediate vector pBI121-M-INT. Two pairs of specific primers with restriction sites were designed to amplify a 351 bp long cDNA conserved sequence fragment of vemalizational gene FaCONSTANS for RT-PCR. After restriction enzyme digestion, the amplified fragment was inserted forwardly and reversely at two sides of the intron of intermediate vector to construct an RNAi expression vector with hairpin structure. [ Result ] Double digestion （HindIII ＋ BamHI） showed that the intron was successfully insert- ed into the vector pBI121. PCR amplification and double digestion indicated that target fragment FaCONSTANS was successfully inserted forwardly and reversely in- to the intermediate vector. [ Conclusion] This study laid foundation for breeding novel flowering-inhibited tall rescue cultivars.

高羊茅FaCONSTANS基因的表达及功能分析

植物由营养生长向生殖生长转变过程中光周期调控起着重要的作用。CONSTANS (CO)是光周期途径中的特有基因,为探讨高羊茅FaCONSTANS (FaCO)基因响应日照长短从而启动植物开花的机理,利用实时荧光定量qRT-PCR技术分析在长日照、短日照、持续光照、持续黑暗条件下FaCO基因的表达水平。构建过表达载体p1300-FaCO,利用农杆菌介导法遗传转化拟南芥,构建沉默载体p1300-FaCO-RNAi遗传转化高羊茅。结果表明,FaCO基因的表达受光周期调控,与生物钟控制的昼夜节律相关。在长日照条件下FaCO基因促进拟南芥开花,且恢复拟南芥突变体开花表型。RNAi沉默FaCO基因的高羊茅转基因植株晚花或者一直处于营养生长阶段。本研究初步探究高羊茅FaCO基因对开花过程的调控,这将有助于更进一步了解该基因的生物学功能。