一株动性球菌对苜蓿的促生和缓解NaHCO_3胁迫作用

为改良和利用盐碱地,以大庆草原盐碱土为材料,筛选耐盐碱细菌,对筛选纯化后的细菌用16S r RNA基因序列对其进行分子生物学鉴定,并对其能力进行分析;设置2种处理(CK1:无菌蒸馏水;S1:菌液),对苜蓿种子进行发芽实验,比较不同处理下种子的发芽率、苗长和鲜重;同样,设置2种处理(CK2:无菌蒸馏水;S2:菌液),采用蛭石浇灌营养液的培养方式进行试验,分析不同处理下的NaHCO_3胁迫苜蓿生长特性及其抗氧化活性变化。结果表明,实验共分离获得43株菌,其中,一株编号为38的菌株(S38),基于16S rRNA基因序列鉴定发现,该菌为动性球菌(Planococcus)属的菌株,具有固氮和产ACC脱氨酶的能力;S1相比CK1处理,显著提高了苜蓿种子的萌发率、苗长和鲜重;S2相比CK2处理,地上部和地下部生物量分别提高32%和50%,Chla、Chlb和Chla+Chlb含量分别增加了1.33、1.15和1.28倍,而可溶性糖含量、抗氧化酶(SOD、POD、CAT)活性均有所降低,同时也影响了K+吸收和分布。由此可见,S38对苜蓿种子有促生的作用,可影响NaCO_3胁迫下苜蓿幼苗生长形态、根形态建成、光合作用等,降低苜蓿对盐的吸收,从而促进幼苗生长。综上,S38在缓解NaHCO_3对植物的伤害方面有着重要的作用。

Over-Expression of ICE1 Gene in Transgenic Rice Improves Cold Tolerance

ICE1, an Arabidopsis thaliana transcription factor gene, was cloned by RT-PCR and successfully transformed into rice variety Kenjiandao 10 by the Agrobacterium-mediated transformation method. PCR amplification and Southern blot analysis indicated that ICE1 had been integrated into rice genome. Compared with the non-transgenic plants, the transgenic plants exhibited high resistance to hygromycin B and were consistent with the Mendelian inheritance of a single copy of the transgenic ICE1. Under the low temperature stress, the transgenic plants showed the lower mortality rate and the increased proline content. These results suggest that the Arabidopsis ICE1 is functional in rice and the over-expression of ICE1 improves the tolerance to cold stress in rice.

Comparative Transcriptome Profiling of Glycine soja Roots Under Salinity and Alkalinity Stresses Using RNA-seq

Saline-alkaline stress can dramatically inhibit plant growth and limit crop production. Wild soybean （Glycine soja） is a crop that adapts well to such environmental stresses. In this study, RNA-sequencing technology was used to analyze the transcriptome profles of G. soja roots subjected to 50 mmol·L^-1 NaHCO3 and 150 mmol·L^-1 NaCl treatments. Totally, 2 125 differentially-expressed genes （DEGs） after NaCl treatment and 1 839 DEGs after NaHCO3 treatment were identifed. The top 14 DEGs revealed by RNA-seq were analyzed using qRT-PCR （quantitative real-time polymerase chain reaction）. Gene ontology （GO） annotation showed that most of DEGs under salt and alkali stresses were enriched in ＂metabolic process＂, ＂catalytic activity＂ and ＂binding＂ terms. To search for transcription factors （TFs） among DEGs, the data were screened against TF database PlantTFDB, and it was found that fve TF families, Apetala2/ethylene-responsive element binding proteins （AP2-EREBP）, V-myb avian myeloblastosis viral oncogene homolog （MYB）, WRKYGQK and Zinc fnger motif （WRKY）, NAM, ATAF1/2, CUC1/2 （NAC） and Cys2/His2 （C2H2） were involved in salt stress response. Other fve TF families, NAC, WRKY, MYB, AP2-EREBP and bZIP were involved in response to alkali stress. These two stress treatments shared NAC, WRKY, AP2-EREBP and MYB, and the only two different TFs were bZIP and C2H2. Forty-eight MYB TFs were differentially expressed under salt and alkali stresses, and most of them were up-regulated. This study provided useful information for further investigation of DEGs and TFs in response to saline and alkaline stresses and helped in understanding the molecular basis of the response of G. soja to saline and alkaline stresses.

盐单胞菌DSM 16354^(T)中新型耐盐基因的克隆及解析

目的:嗜盐菌分布广泛且适应能力极强,为加强对嗜盐菌耐盐机制的探索与研究,从盐单胞菌Halomonas alkaliphila DSM 16354^(T)中筛选出潜在的与耐盐有关的基因,对其进行生物信息学分析,并验证相关蛋白的生理功能。方法:利用基因文库筛选和功能互补相结合的方法,通过与大肠杆菌(Escherichia coli)盐敏感缺陷株KNabc(ΔnhaA、ΔnhaB、ΔchaA)的耐盐功能互补实验,筛选出具有耐盐功能的蛋白编码基因,并通过荧光猝灭恢复实验测定蛋白的逆向转运活性以及底物亲和力。结果:筛选得到两个具有耐盐功能的蛋白编码基因,生物信息学分析结果表明该基因编码来自于DUF1538(domain of unknown function with No.1538 family)家族功能未知的膜蛋白,分别命名为duf1和duf2。系统发育树分析结果表明,来自盐单胞菌DSM 16354^(T)中的DUF1、DUF2属于一个独立的分支,预测这两个蛋白可能是DUF1538家族转运蛋白的新成员。对DUF1和DUF2的生理功能进行分析,发现duf1和duf2单独表达时均不具有耐盐碱能力,而共同表达时则表现出显著的耐盐碱功能,表明DUF1和DUF2两个亚基共同支持了蛋白的耐盐碱功能。蛋白的逆向转运测定活性结果表明双组份蛋白DUF1-2具有Na^(+)(Li^(+)、K^(+))/H^(+)逆向转运活性。结论:筛选得到的基因duf1和duf2共同表达时具有盐碱耐受功能以及逆向转运蛋白活性,这为筛选出新的DUF1538家族转运蛋白基因和进一步探究DUF1538家族转运蛋白功能奠定了基础。