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.     

东北次生林森林公园林内景观质量提升技术

为提高东北次生林森林公园景观质量,在吉林市大石门沟森林公园次生林选择典型样地,采取了修枝、去除枯落物、清灌等技术措施对次生林林内进行处理。采用心理物理学派的美景度评价法(SBE)、相关分析和回归分析等方法,确定了次生林样地内景观质量指标和预测模型。结果表明:3种技术措施均能有效提高林内景观质量,清灌措施使林内景观美景度均值从29.8提升到70.3;修枝措施使林内景观美景度均值从38.3提升到58.3;去除枯落物措施使林内景观美景度均值从36.9提升到53.4。对照组美景度的模型如下:M_(SBE)=-49.7+31.2X_(10),空间感(X_(10))为主导因子(P0.01);清灌组的美景度回归模型:M_(SBE)=-132+56.5X_4,自然程度(X_4)为主导因子(P0.05);修枝组的美景度回归模型:M_(SBE)=-102.5+58.1X_5,树木排列(X_5)为主导因子(P0.01);去除枯落物组的美景度回归模型:M_(SBE)=-73.2+39.6X_1,树干形态(X_1)为主导因子(P0.05)。说明3种措施中提升景观质量效果排序,清灌修枝去除枯落物;采取不同的技术措施使原有次生林林内景观结构发生改变,影响景观美景度的主要因子也同时发生的改变。     

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。     

国内外应对气候变化立法研究进展

为了积极应对气候变化,促进低碳经济发展,分析了当前应对气候变化立法方面的研究进展,认为尽管取得了一定进展,但现有法律既不能满足我国气候外交的需求,也不足以担当气候治理的重任,我国还缺乏系统性专门性的应对气候变化法律.为了应对气候变化的挑战,必须充分整合国内外应对气候变化立法方面的科技资源,以保证我国气候外交和气候治理战略的科学性.     

Cloning of Buffalo BMP1 Gene and its Expression Pattern Investigation in Different Tissues

Buffalo BMP1 gene was cloned in the present study, the BMP1 sequence was systemically analysed by bioinformatics techniques, and the expression level of BMP1 gene in different tissues were also assayed with Real-time fluorescence quantitative PCR (QRT-PCR).The results showed that with RT-PCR a 3 195 bp buffalo BMP1 gene was cloned and sequenced, including the whole ORF of 2 967 bp (coding 988 amino acid).The sequence multialigned results showed that buffalo BMP1 gene shared 99%, 96%, 96%, 96% and 95% of similar amino acid sequence with Bos taurus, Sus scrofa, Equus, Homa sapiens and Mus musclus, respectively.It was predicted that buffalo BMP1 protein contained a signal peptide domain, a preregion, a metalloproteinases domain, five complement-Uegf-BMP-1 domain (CUB domain) and two epitheloid growth factor-like domain (EGF-like domain).In addition, we also analyzed the expression level in different tissues through QRT-PCR, the results showed that BMP1 gene mRNA existed in all nine tissues with the most abundant expression in heart, followed by testis, ovary, genital ridge, while with lower amount of bone and other tissues, the minimal expression in liver was observed.     

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。