高通量转录组RNA-seq研究成年牛下丘脑-垂体-卵巢

肉牛的性成熟受下丘脑-垂体-卵巢(HPO)轴的调节,但其具体作用机制尚不清楚.为了探讨HPO轴繁殖相关基因和调控机制,采用高通量转录组RNA-seq技术,研究了成年安格斯牛(30个月大)的下丘脑、垂体和卵巢.结果表明,HPO共有18 179个基因在GO(基因本体论)中注释,37 158个基因在KEGG(京都基因百科全书和基因组)中注释,36 645个基因在KOG(KEGG同源数据库)中注释.GO分析表明,HPO富集了8种生殖相关功能,主要是受精、单个组织生殖过程、子宫胚胎发育和雌激素反应.KEGG分析发现,HPO富集300条信号通路(SP),主要是癌症SP和磷脂酰肌醇3激酶蛋白激酶(PI3K-Akt)SP.与生殖有关的SPs共有16条,主要是Notch SP、神经营养素SP、哺乳动物雷帕霉素(mTOR)SP和血管内皮生长因子(VEG F)SP.为今后肉牛繁殖性能的提高、分子育种和同行参考提供了科学依据.     

Congruent spatial patterns of species richness and phylogenetic diversity in karst flora: Case study of Primulina (Gesneriaceae)

The karst landform in southern China is renowned for its high levels of species diversity and endemism. Globally, karst ecosystems are under threat from unsustainable anthropogenic disturbance and climate changes and are among the most threatened ecosystems worldwide. In this study, we used the typical karst endemic genus in southern China, Primulina Hance, as a model to identify areas within the karst landform with high diversity and to investigate congruence between phylogenetic and species‐based measures of diversity. Using phylogenetic information and species distribution data, we measured geographical patterns of diversity with four metrics: species richness (SR), corrected weighted endemism (CWE), phylogenetic diversity (PD), and phylogenetic endemism (PE). Our results revealed a high spatial congruence among SR, PD, and PE, with hotspot areas identified in the Nanling Mountains (i.e., north Guangdong and northeast Guangxi) and southeast Yungui Plateau (i.e., north and southwest Guangxi), whereas the hotspots of CWE are comparatively uniform throughout the geographic extent. The categorical analysis of neo‐ and paleoendemism identified a pattern of mixed neo‐ and paleoendemism in numerous grid cells, suggesting that karst areas in southern China have acted as both “museums” and “cradles” of plant evolution. Conservation gap analysis of hotspots revealed that the majority of prioritized hotspots (>90%) of the genus are outside of protected areas, therefore indicating the limited effectiveness of national nature reserves for the karst flora. Overall, our results suggest that the karst flora merits more conservation attention and SR can be an effective surrogate to capture PD in conservation planning.     

Evolution and diversity of axon guidance Robo receptor family genes

The diversity of axon guidance (AG) receptors reflects gains in complexity of the animal nervous system during evolution. Members of the Roundabout (Robo) family of receptors interact with Slit proteins and play important roles in many developmental processes, including AG and neural crest cell migration. There are four members of the Robo gene family. However, the evolutionary history of Robo family genes remain obscure. We analyzed the distribution of Robo family members in metazoan species ranging in complexity from hydras to humans. We undertook a phylogenetic analysis in metazoans, synteny analysis, and ancestral chromosome mapping in vertebrates, and detected selection pressure and functional divergence among four mammalian Robo paralogs. Based on our analysis, we proposed that the ancestral Robo gene could have undergone a tandem duplication in the vertebrate ancestor; then one round of whole genome duplication events occurred before the divergence of ancestral lamprey and gnathostome, generating four paralogs in early vertebrates. Robo4 paralog underwent segmental loss in the following evolutionary process. Our results showed that Robo3 paralog is under more powerful purifying selection pressure compared with other three paralogs, which could correlate with its unique expression pattern and function. Furthermore, we found four sites under positive selection pressure on the Ig1‐2 domains of Robo4 that might interfere with its binding to Slits ligand. Diverge analysis at the amino acid level showed that Robo4 paralog have relatively greater functional diversifications than other Robo paralogs. This coincides with the fact that Robo4 predominantly functions in vascular endothelial cells but not the nervous system.     

盐增强培养对弗氏链霉菌产新霉素的影响

目的: 弗氏链霉菌(Streptomyces fradiae)作为氨基糖苷类抗生素新霉素的主要生产菌株,其新霉素B具有抗菌活性强、抗癌、抗HIV等作用,提高新霉素B的效价具有重要意义。方法: 在满足微生物正常生长所需盐离子的条件下,通过盐增强培养的方式向培养基中添加不同种类、浓度无机盐来改变细胞壁附近的理化特性、渗透压以及培养基中的碳氮比。结果: 不同无机盐对新霉素B效价影响程度由高到低为(NH₄)₂SO₄、NaCl、KCl、K₂SO₄;当添加60 mmol/L(NH₄)₂SO₄时新霉素B的效价达到最高为15 864 U/mL,而NaCl在80 mmol/L浓度时产量最高为7 429.7 U/mL,相比未添加无机盐分别提高3.8倍和0.82倍。在含有80 mmol/L的NaCl的发酵培养基中添加不同浓度的(NH₄)₂SO₄并定时取样检测其中氨基氮、还原糖、pH、TG、菌浓以及新霉素B效价的变化,发现60 mmol/L(NH₄)₂SO₄浓度下氨基氮、还原糖、TG的消耗速率加快,菌体比生长速率μ最大为0.097/h,新霉素B的合成加快且产量提高为17 399 U/mL,同时菌丝呈现出聚集变短的形态且产孢提前。结论: 盐增强培养方式既可以作为一种形态学工程手段,通过改变弗氏链霉菌的微观形态来促进次级代谢产物新霉素B产量的提高,同时也可以作为培养基优化策略来提高新霉素B的产量,这为进一步提高弗氏链霉菌中次级代谢产物产量的研究奠定了基础。     

冬虫夏草菌实时荧光定量PCR内参基因的筛选

以冬虫夏草单子囊孢子分离得到的菌株TZ8-1的3种菌丝形态为实验材料,提取RNA,经反转录获取cDNA,选择了11个持家基因为候选内参基因（18S rRNA、APRTase、β-TUB、RPL2、EF1-α、PGI、PGM、H⁺-ATPase、ACT1、UBQ和GAPDH）,根据该菌基因组注释结果来设计引物,采用实时荧光定量PCR（qRT-PCR）技术进行定量扩增,利用geNorm、NormFinder和BestKeeper算法程序进行表达稳定性评估,并用RefFinder对评估结果进行综合排比,最终筛选得到了最适内参基因。结果表明,所选取的11个候选内参基因均可作为冬虫夏草菌菌丝体时期的内参基因,稳定性最好的3个内参基因分别是UBQ、PGE和ACT1,稳定性较差的3个内参基因分别是GAPDH、RPL2和β-TUB。     

Benefiting others and self: Production of vitamins in plants

Vitamins maintain growth and development in humans, animals, and plants. Because plants serve as essential producers of vitamins, increasing the vitamin contents in plants has become a goal of crop breeding worldwide. Here, we begin with a summary of the functions of vitamins. We then review the achievements to date in elucidating the molecular mechanisms underlying how vitamins are synthesized, transported, and regulated in plants. We also stress the exploration of variation in vitamins by the use of forward genetic approaches, such as quantitative trait locus mapping and genome-wide association studies. Overall, we conclude that exploring the diversity of vitamins could provide new insights into plant metabolism and crop breeding.     

A chloride efflux transporter,BIG RICE GRAIN 1, is involved in mediating grain size and salt tolerance in rice

Grain size is determined by the size and number of cells in the grain. The regulation of grain size is crucial for improving crop yield; however, the genes and molecular mechanisms that control grain size remain elusive. Here, we report that a member of the detoxification efflux carrier /Multidrug and Toxic Compound Extrusion (DTX/MATE) family transporters, BIG RICE GRAIN 1 (BIRG1), negatively influences grain size in rice (Oryza sativa L.). BIRG1 is highly expressed in reproductive organs and roots. In birg1 grain, the outer parenchyma layer cells of spikelet hulls are larger than in wild-type (WT) grains, but the cell number is unaltered. When expressed in Xenopus laevis oocytes, BIRG1 exhibits chloride efflux activity. Consistent with this role of BIRG1, the birg1 mutant shows reduced tolerance to salt stress at a toxic chloride level. Moreover, grains from birg1 plants contain a higher level of chloride than those of WT plants when grown under normal paddy field conditions, and the roots of birg1 accumulate more chloride than those of WT under saline conditions. Collectively, the data suggest that BIRG1 in rice functions as a chloride efflux transporter that is involved in mediating grain size and salt tolerance by controlling chloride homeostasis.     

MiR319-targeted OsTCP21 and OsGAmyb regulate tillering and grain yield in rice

Multiple genes and microRNAs (miRNAs) improve grain yield by promoting tillering. MiR319s are known to regulate several aspects of plant development; however, whether miR319s are essential for tillering regulation remains unclear. Here, we report that miR319 is highly expressed in the basal part of rice plant at different development stages. The miR319 knockdown line Short Tandem Target Mimic 319 (STTM319) showed higher tiller bud length in seedlings under low nitrogen (N) condition and higher tiller bud number under high N condition compared with the miR319a-overexpression line. Through targets prediction, we identified OsTCP21 and OsGAmyb as downstream targets of miR319. Moreover, OsTCP21 and OsGAmyb overexpression lines and STTM319 had increased tiller bud length and biomass, whereas both were decreased in OsTCP21 and OsGAmyb knockout lines and OE319a. These data suggest that miR319 regulates rice tiller bud development and tillering through targeting OsTCP21 and OsGAmyb. Notably, the tiller number and grain yield increased in STTM319 and overexpression lines of OsTCP21 and OsGAmyb but decreased in OE319a and knockout lines of OsTCP21 and OsGAmyb. Taken together, our findings indicate that miR319s negatively affect tiller number and grain yield by targeting OsTCP21 and OsGAmyb, revealing a novel function for miR319 in rice.