CRISPR系统促进脂肪干细胞成骨分化与抑制成脂分化的研究

本研究旨在优化组织培养法分离小鼠脂肪间充质干细胞（adipose-derived stem cells,ASCs）,为研究成骨分化和成脂分化在间充质干细胞分化过程中的相互影响奠定基础。通过细胞形态学观察、细胞生长曲线和流式仪器检测所分离获得的间充质干细胞的特性,利用CRISPR-dCas9系统在快速促进间充质干细胞成骨分化的前提下观察其对成脂分化的影响,并通过生化染色、实时荧光定量PCR和免疫细胞学等手段进行分析。结果显示,接种3~5 d后可见细胞从组织块周围爬出,光镜下可见细胞形态多为成纤维细胞样的梭形细胞,且形态单一均匀,具有较高的爬出率,可以大大提高脂肪间充质干细胞的分离效率;通过CRISPR-dCas9系统激活Runx2和Osterix基因后可以促进间充质干细胞的成骨分化,实时荧光定量PCR及油红O染色结果显示,CRISPR-dCas9系统可以同时抑制间充质干细胞的成脂分化;通过CRISPR-dCas9-KRAB系统同时抑制成骨相关基因Runx2和Osterix后可以促进成脂分化。本研究利用组织贴壁法成功获得了高纯度的脂肪间充质干细胞,具有间充质干细胞的特性和分化能力;利用CRISPR系统可以同时过表达Runx2和Osterix两个基因,可以在进成骨分化的同时抑制成脂分化,表明成脂分化和成骨分化的相关性,为基因编辑在间充质干细胞诱导分化和临床应用方面提供了新的思路和方法。     

当前我国农村土地制度改革的必要性与对策思考

我国农村土地制度与"三农"问题紧密相关。与其他国家的农村土地制度相比,我国的农村土地制度更具独特性,因此推进我国农村土地制度改革必须立足基本国情,而不能照抄他国的土地制度。该文分析了当前我国农村土地制度的必要性,并对推进我国农村土地制度改革提出对策思考。     

葡萄霜霉菌候选效应子RXLR5信号肽的鉴定

利用Getorf、SignalP 3.0、BLASTP等生物信息学软件和PERL语言从葡萄霜霉菌基因组中预测到一条编码RXLR-EER结构域的效应蛋白候选基因,将该基因编码的蛋白命名为RXLR5。将RXLR5信号肽SP5编码序列连接到pSUC2T7M13ORI载体并转化到酵母蔗糖酶缺陷菌株YTK12后,重组菌株能成功分泌蔗糖酶,促使棉籽糖分解成单糖,因此,能在YPRAA培养基上正常生长;同时生成的单糖能与TTC反应产生红色不溶于水的氯化三苯基四氮唑。由此推测,SP5具有信号肽活性,可能在RXLR5从葡萄霜霉菌细胞内泌到胞外的过程中起重要作用。     

Seasonal changes in the crude protein concentration of mixed swards of white clover/perennial ryegrass grown without fertilizer N in an organic farming system in the United Kingdom

Changes in the crude protein (CP) concentration of white clover and perennial ryegrass herbage from a mixed sward were determined on six sampling dates from May to October in each of 2 years. The swards were grown without fertilizer N in an organic farming system and continuously grazed by dairy cows during the grazing season. The annual mean contents of white clover in the dry matter (DM) of the sward were 272·3 and 307·0 g kg⁻¹ in Years 1 and 2. The mean CP concentrations of the white clover and perennial ryegrass herbage were 251·6 and 151·9 g kg⁻¹ DM in Year 1 and 271·9 and 174·0 g kg⁻¹ DM in Year 2 respectively. The CP concentration of the white clover increased significantly during the grazing season from 220·0 to 284·1 g kg⁻¹ DM in Year 1 and from 269·0 to 315·5 g kg⁻¹ DM in Year 2. In the perennial ryegrass herbage the CP concentration increased from 112·2 to 172·6 g kg⁻¹ DM in Year 1 and from 142·7 to 239·5 g kg⁻¹ DM in Year 2. The rate of increase during the season in the CP concentration of the perennial ryegrass herbage was similar to the rate of increase recorded in the white clover herbage.     

Improved milk production through PG–PL system by provision of in‐house shelter management in lactating Murrah buffaloes during winter season

Comprehensive information on the role of β‐casein and plasminogen–plasmin (PG–PL) system in milk secretion of Murrah buffaloes during winter season is lacking, although effects of cold stress can be ameliorated to an extent by altering microclimate at farm level. Hence, this study was aimed to determine the changes in productivity along with PG–PL system of milk, plasma hormones and metabolites of buffaloes during winter (December–January) season under two different management systems. Average minimum temperature and wind chill index during this season were 7.02 and 12.74 °C respectively. Buffaloes were divided in two groups of six animals each: control and treatment, where treatment group animals were placed in‐house with floor bedding of paddy straw and the control group animals in loose housing system without straw bedding. Physiological responses were recorded, and milk and blood samples were collected at weekly intervals for six‐week experimental period. Under in‐house management system, buffaloes experienced better comfort by alleviating environmental stress as their physiological responses such as respiration rate and pulse rate were significantly reduced (p < 0.01) as compared to the control, which subsequently resulted higher milk yield by 9.92% (p < 0.05). Analysis of milk samples revealed higher concentration of plasminogen (10.6 vs. 8.05 μg/ml; p < 0.01) and β‐casein (p < 0.05), and lower plasmin level (0.299 vs. 0.321 μg/ml; p < 0.05) in buffaloes under treatment group. It was also found that plasma cortisol, glucose and non‐esterified fatty acids levels were higher (p < 0.01) in control group as compared to the treatment animals by 13.6%, 8.14% and 12.6% respectively. However, milk composition, growth hormone, epinephrine and norepinephrine level in plasma were similar in both the groups. Hence, it may be concluded that provision of in‐house shelter management with floor bedding of paddy straw during winter was effective to minimize environmental stress and improved milk production through manipulation of PG–PL system in buffaloes.     

苜蓿-禾草混播对根系可溶性糖含量的影响

可溶性糖对植物的抗冻性存在着积极的影响。为研究苜蓿（Medicago sativa）与禾草混播对根系可溶性糖含量的影响,以苜蓿与无芒雀麦（Bromus inermis Leyss）、披碱草（Elymus dahuricus Turcz）、虉草（Phalaris arundinacea L.）按豆禾间行比例1∶1(B1)、2∶2(B2)、1∶2(B3)和2∶1(B4)混播,并在初霜后测定不同混播处理牧草根系的可溶性糖含量。结果表明,禾草种类对苜蓿根系可溶性糖含量的影响不显著;豆禾间行比例2∶2混播处理苜蓿根系可溶性糖含量显著低于1∶2和2∶1混播处理,1∶1、1∶2和2∶1混播处理间无显著差异;苜蓿与虉草混播后,不同混播比例的虉草根系可溶性糖含量都高于单播虉草。3种禾草中,虉草根系可溶性糖含量显著高于其余两种。综上所述,豆禾间行比为1∶1及1∶2混播处理中苜蓿及禾草的根系可溶性糖含量均较高,考虑苜蓿的耐寒力往往弱于3种禾草,又以间行比1∶2混播的处理为最佳选择。研究为当地苜蓿与禾草混播草地的持续利用提供理论参考。     

中国矮生刺黄瓜品系特性的研究初报

本文利用由中国蔓生刺黄瓜自然突变产生的矮生型黄瓜为材料，通过多低自交选育及性状观察，初步了解该矮生黄瓜植物学性状除具有自封顶、植株矮化性状外，与蔓生型黄瓜无明显差异。在生物学方面，该矮生黄瓜具有早熟、雌花节率高、结果集中、生长期短等特性。通过对中国矮生刺黄瓜与不同品种蔓生黄瓜杂交后代的观察、分析，初步认为黄瓜植株高度是由单基因体系和多基因体系共同作用决定的。     

Modelling land use strategies to optimise crop production and protection of ecologically important weed species

There is a need to develop farming systems that enable both a satisfactory level of crop production and suitable environmental conditions for natural species. Wildlife‐friendly cropping techniques, such as a reduced amount of applied herbicide or a lower crop density, might be adopted in order to maintain populations of weed species of biological interest. An alternative might be to adopt an intensive cropping system in a part of the field and spare the other part as set‐aside or field margins, available for the natural development of plant species. The objective of this study was to present a method to compare two strategies for maintaining a desirable level of abundance of a given species of interest in agricultural areas, specifically (i) a strategy based on a wildlife‐friendly cropping system in a large cultivated area and (ii) a strategy based on a more intensive cropping system in a reduced area of cultivation, i.e. with land‐sparing. The principle is to calculate the ratio of crop production obtained with strategy (i) to the production obtained with strategy (ii) for a given target density of natural species. We show that the value of this ratio, and thus the relative performance of the two strategies, depends on the density of the weed species that can be maintained in an uncultivated ecological area. The method is applied in case studies of two plant species with contrasting ecology and conservation goals. The numerical results show that the strategy based on a wildlife‐friendly cropping system is more profitable in most situations.     

Histopathological Changes in Third-Instar and Adult Anastrepha ludens (Diptera: Tephritidae) After in vitro Heat Treatment

The Mexican fruit fly, Anastrepha ludens Loew (Diptera: Tephritidae), is one of the most harmful pests of mango causing direct damage by oviposition on the fruit pulp. Mango for export is subjected to hydrothermal treatment as a quarantine method for the control of this pest, but exposure to heat for long periods of time reduces considerably the quality and shelf-life of treated fruit. The aim of this work was to study morphological changes of third-instar larvae and adults of A. ludens after in vitro exposure to high temperature at sublethal times. A heating block system was used to expose larvae at 46.1°C for 19.6 and 12.9 min, producing 94.6 and 70% mortality, respectively. Treated larvae were processed for optical microscopy. A fraction of surviving treated larvae was separated into containers with artificial diet to allow development into adults. Adult sexual organs were dissected and processed for transmission electron microscopy analysis. Results showed that 94.6% of the treated larvae died at 46.1°C for 19.6 min and none of the surviving larvae eclosed to adulthood, as they developed as malformed puparia. For the in vitro treatment at 46.1°C during 12.9 min, 70% of the treated larvae died and only 3.75% reached the adult stage, but ultrastructural damage in the male testes and in the female ovaries was observed. Additionally, 11.1% of the adult flies from the in vitro treatment also showed wing malformation and were incapable of flying. The analysis showed that surviving flies were unable to reproduce.     

利用Ph基因诱导小麦外源染色体易位

对小麦染色体配对控制体系的调控进行了综述,即利用5B染色体缺失效应、Ph基因突变系和Ph抑制基因等方法来诱导易位系。该方法产生的易位系遗传补偿性较好。