Improvements of TKC Technology Accelerate Isolation of Transgene-Free CRISPR/Cas9-Edited Rice Plants

Elimination of the CRISPR/Cas9 constructs in edited plants is a prerequisite for assessing genetic stability, conducting phenotypic characterization, and applying for commercialization of the plants. However, removal of the CRISPR/Cas9 transgenes by genetic segregation and by backcross is laborious and time consuming. We previously reported the development of the transgene killer CRISPR (TKC) technology that uses a pair of suicide genes to trigger self-elimination of the transgenes without compromising gene editing efficiency. The TKC technology enables isolation of transgene-free CRISPR-edited plants within a single generation, greatly accelerating crop improvements. Here, we presented two new TKC vectors that show great efficiency in both editing the target gene and in undergoing self-elimination of the transgenes. The new vectors replaced the CaMV35S promoter used in our previous TKC vector with two rice promoters to drive one of the suicide genes, providing advantages over our previous TKC vector under certain conditions. The vectors reported here offered more options and flexibility to conduct gene editing experiments in rice.     

Identification of Differentially Expressed Genes and Lipid Metabolism Signaling Pathways between Muscle and Fat Tissues in Broiler Chickens

In this study, signaling pathways and key differentially expressed genes (DEGs) involved in lipid metabolism in muscle and fat tissues were investigated. Muscle and abdominal fat tissues were obtained from 35-day-old female broilers for RNA sequencing. DEGs between muscle and fat tissues were identified. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses of DEGs were performed. A total of 6130 DEGs were identified to be significantly enriched in 365 GO terms, most of which were involved in biological processes, cellular components, and molecular functions in muscle and fat tissues. Three important lipid signaling pathways (pyruvate metabolism, the insulin signaling pathway, and the adipocytokine signaling pathway) were identified among the fat and muscle tissues of broilers. The key common DEGs in these pathways included phosphoenolpyruvate carboxykinase 2 (PCK2), acetyl-CoA carboxylase 1 alpha and beta (ACACA and ACACB), and the mitogen-activated protein kinase (AMPK) gene family. Hence, our findings revealed the pathways and key genes and gene families involved in the regulation of fat deposition in the muscle and fat tissues of broilers.     

7株根瘤菌菌株与热研2号柱花草共生最适磷浓度研究

采用水培的方法,设3个磷水平,7个菌株,通过对株高、茎叶鲜重、茎叶干重、固氮酶活性、茎叶氮含量、茎叶磷含量的测定及综合分析,确定使热研2号柱花草接种不同根瘤菌菌株达到最佳促生性能的磷浓度。结果表明:7个菌株在中磷时鲜重和含氮量最大,低磷不利于鲜重和含氮量的增加;经主因子综合分析,菌株YM11-1、RJS9-2、PN13-3在高磷时对热研2号柱花草有最好的固氮促生效果,菌株LZ3-2、BS1-1、PN13-3在中等磷条件下有最好的固氮促生效果,菌株FS3-1-1在高磷和中磷都有最好的固氮促生效果。     

Dvl1及其突变体腺病毒载体的构建与在MSCs中表达鉴定

Dvl1及其结构缺失突变体ΔDIX（dsh and axin）和ΔDEP（dsh,Egl-10 and pleckstrin）腺病毒载体的构建并验证其在MSCs（mesenchymal stem cells）中的表达情况。将目的基因定向克隆至pAdTrack-CMV穿梭载体上,并经PmeⅠ线性化后在BJ5183细菌中与pAdEasy-1骨架质粒同源重组,获得重组腺病毒载体,在QBI-293A细胞中包装及扩增,实时荧光定量PCR和Western bolt验证Dvl1在MSCs中的表达情况。经PCR、PacⅠ单酶切鉴定及测序分析,成功构建Ad-Dvl1、Ad-ΔDIX和Ad-ΔDEP腺病毒载体,目的基因序列与GenBank报道一致,并选出150 MOI为最适感染MSCs的感染复数,成功构建了Ad-Dvl1、Ad-ΔDIX和Ad-ΔDEP腺病毒载体,并获得高滴度的病毒子,qPCR和Western blot证实Dvl1在MSCs中高表达并增加了β-catenin在细胞中的累积,为进一步研究Dvl1在MSCs迁移过程的作用奠定了基础。     

Symbiotic N<Subscript>2</Subscript> fixation and nitrate utilisation in irrigated lucerne (<Emphasis Type="Italic">Medicago sativa</Emphasis>) systems

Symbiotic N₂ fixation by lucerne (Medicago sativa) has capacity to provide significant inputs of N to agro-ecosystems, and the species has also been shown to scavenge soil mineral N and thus act as a sink for excess reactive N. The balance between these two N cycle processes was investigated in an extensive irrigated lucerne growing region where nitrate contamination of groundwater has been reported. We sampled 18 permanent pure lucerne stands under irrigation for standing dry matter, total shoot N, and N₂ fixation using ¹⁵N natural abundance along with activity of the inducible enzyme nitrate reductase as indicators of use of soil NO₃⁻ by lucerne. On average 65% of lucerne N was obtained from symbiotic N₂ fixation. Converting standing dry matter estimates to annual N₂ fixation amounts we calculated average N₂ fixation of 311 kg N/ha, including N in roots and nodules. Uptake of N from soil by lucerne was calculated to be 181 kg N/ha/year. We were not able to identify the source of this soil mineral N, although nitrate reductase activity of lucerne was higher than that of non-N₂ fixing species examined.     

京郊边际土地纤维素类能源草规模化种植与管理技术

为探索能源植物在北京及周边地区的发展潜力,在京郊挖沙废弃地、荒滩地、撂荒地、污染农田4种边际土地上开展了柳枝稷、荻、芦竹和杂交狼尾草4种纤维素类能源草的规模化种植。在此基础上,总结提出了京郊边际土地纤维素类能源草规模化种植与管理技术,为基于边际土地发展京郊生物质原料生产与生物质能源产业奠定了良好的技术基础。     

巴西橡胶树乳管细胞HbWRKY1基因克隆及表达分析

采用PCR和RACE技术,从巴西橡胶树无性系热研7-33-97的乳管细胞中克隆了WRKY转录因子家族的一个成员HbWRKY1。该基因全长为1755 bp,含有1个1440 bp阅读框,编码479个氨基酸。生物信息学分析表明,HbWRKY1含有2个WRKY结构域和C2H2锌指结构基序,与蓖麻、杨树、葡萄、黄瓜、拟南芥的WRKY成员的氨基酸序列同源性分别为64.14%、60.04%、42.54%、40.61%和35.4%,属于Ⅰ类WRKY家族成员。实时荧光定量PCR分析表明,割胶和乙烯显著上调HbWRKY1基因的表达,但茉莉酸和机械伤害对HbWRKY1的表达没有明显影响,表明HbWRKY1可能在乙烯信号途径对防卫蛋白的调节中起重要作用。     

环植所科研主体向海口转移工作与方案研究

中国热带农业科学院环境与植物保护研究所是从事热带农业环境与植物保护领域研究的国家级公益类非营利性研究所。海口环境与植物保护研究实验室项目工程即将竣工，该所科研主体由儋州向海口转移工作已列入所2011年计划的第一项重点工作。从工作方案入手，重点从领导管理、群众参与、宣传引导、督导落实等方面提出建议确保转移工作平稳有序。     

马克思主义生态思想的实践视阈 ——对促进安徽省旅游业可持续发展的启示

马克思恩格斯生态思想是建设社会主义生态文明的理论渊源和行动指南，对于促进资源节约型、环境友好型社会建设有着重要的指导意义。在安徽省旅游业已经进入了转型升级的关键时期，返本开新，深入挖掘马克思主义生态思想的时代精华，对于我们贯彻科学发展观，促进旅游业的可续、健康、快速发展有着不可或缺的启迪作用。