DNA去甲基化参与提高白菜型冬油菜抗寒性的生理机制

利用来自不同育种环境的10份白菜型冬油菜材料,通过半致死温度的测定分析其抗寒性与环境的关系,并利用不同浓度DNA甲基化抑制剂5-azaC处理,分析DNA去甲基化对白菜型冬油菜DNA整体甲基化水平及低温胁迫下苗期生理特性的影响。结果表明,不同育种环境选育的材料,其半致死温度具有显著差异,其中材料2018-FJT、DT-7、DT-9与MXW-1的半致死温度分别为-16.04℃、-15.98℃、-15.63℃、-15.04℃;材料CT-2360、CT-2380、CT-2400、CT-2420、CT-2440、CT-2460的半致死温度分别为-11.32℃、-11.6℃、-11.42℃、-11.44℃、-12.97℃、-13.28℃。依据半致死温度,10个白菜型冬油菜抗寒性由强到弱依次为:2018-FJTDT-7DT-9MXW-1CT-2460CT-2400CT-2380CT-2420CT-2440CT-2360。抗寒性的形成与育成环境的纬度和年平均气温呈极显著正相关。选择抗寒性及产地不同的4个材料CT-2360、MXW-1、2018-FJT、DT-7进行生理测定,结果表明,1 000μmol·L~(-1) 5-azaC处理能显著抑制幼苗的根生长,4个材料的根长较对照减少93.14%～95.06%;低温下5-azaC处理对抗寒性较弱材料CT-2360幼苗的相对电导率、丙二醛含量以及强抗寒性材料DT-7幼苗的脯氨酸、可溶性蛋白含量影响最显著(P0.05);低温处理过程中,4个材料的SOD、POD、CAT活性均上升,以低温处理5d最显著(P0.05),其中2018-FJT的SOD活性增幅最高,为45.85%,DT-7的POD活性增幅最大,为460%,CT-2360的CAT活性增幅最显著,为321.02%。HPLC分析发现常温下抗寒性较弱材料CT-2360的甲基化水平为77.48%,高于其他3个抗寒性强的材料;经过5-azaC处理后发生明显的去甲基化作用,证明抗寒能力受DNA去甲基化的调控。     

Nitric oxide impacts bovine sperm capacitation in a cGMP‐dependent and cGMP‐independent manner

We aimed to elucidate whether NO acts in in vitro sperm capacitation in bovine via cGMP/PKG1 pathway. For this, cryopreserved bovine sperm were capacitated in vitro with 20 µg/ml heparin (Control) plus treatments: 1 mM L‐arginine (L‐arg, NO precursor), 50 µM Rp‐8‐Bromo‐β‐phenyl‐1,N²‐ethenoguanosine‐3′,5′‐cyclic monophosphorothioate (Rp‐8‐Br‐cGMPS, selective inhibitor of the binding site for cGMP in PKG1), 1 mM 2‐Phenyl‐4,4,5,5‐tetramethylimidazoline‐1‐oxyl 3‐oxide (PTIO, NO scavenger), and the combinations of L‐arg + RP‐8‐Br‐cGMPS and L‐arg + PTIO. Sperm motility and vigour were determined by phase‐contrast microscopy, capacitation status by chlortetracycline staining, and the intracellular concentration of cGMP was measured by ELISA. Data were subjected to analysis of variance and means compared with SNK test at 5% probability. Motility and vigour were lower in sperm treated with PTIO when compared to Control and other treatments (p < .05). The L‐arg treatment showed the highest percentage of capacitated sperm when compared to the Control and other treatments (Rp‐8‐Br‐cGMPS, L‐arg + Rp‐8‐Br‐cGMPS and PTIO) (69.8 ± 3.4%, 51.2 ± 3.0, 51.1 ± 2.1, 51.2 ± 3.0 and 45.5 ± 2.7, respectively) (p < .05). The capacitation ratio (%) was lower in treatments with Rp‐8‐Br‐cGMPS, L‐arg + Rp‐8‐Br‐cGMPS and PTIO, respectively (p < .05). Lastly, cGMP concentration (pmol/ml) was lower in PTIO and L‐arg + PTIO (1.3 ± 0.3 and 1.6 ± 0.4) and was higher in Rp‐8‐Br‐cGMPS and L‐arg + Rp‐8‐Br‐cGMPS (3.7 ± 0.4 and 4.0 ± 0.5) treatments. We showed that during in vitro capacitation of cattle: (a) NO influences sperm motility and vigour; (b) NO is associated with cGMP synthesis through two independent pathways and (c) the cGMP/PKG1 pathway has a partial role in sperm capacitation and does not involve the L‐arg/NO.     

芸薹种作物抽薹相关基因BrFLC1的CAPS标记

以16份芸薹种作物DH系为试材,通过人工春化处理对其抽薹性状进行了鉴定,明确了材料间抽薹早晚存在显著差异;根据BrFLC1基因序列设计引物进行PCR扩增发现,所有16份材料均获得980 bp的片段。多序列比对与抽薹性的关联分析发现,材料的抽薹早晚与扩增片段的碱基变异相关联,并发现关联位点的碱基变异为限制性内切酶Mva I的识别位点;利用该酶对PCR产物酶切可以将材料明显区分开来,从而建立了BrFLC1基因的CAPS标记（命名为G-Mva I）。利用此标记对96份芸薹种作物DH材料的验证表明,该基因的分子标记结果与抽薹时间表型显著相关,用该标记可以进行分子标记辅助选择。     

施用污泥对黑麦草生育及其元素含量的影响

将兰州市七里河污水处理厂的生活污泥施用于多年生黑麦草,藉以探讨施用量对黑麦草生长及其元素含量的影响。结果表明,随着污泥施用量(0.5～8kg/m²)的增加,黑麦草盖度,密度,地上、地下生物量以及叶绿素含量显著增加。施用后黑麦草地上部组织中N、P、K、Mg、Fe、Zn和Cu的含量增加,Mn含量减少,Ca和重金属元素Hg、Ni、Pb、As含量与对照间差异不显著,没有对黑麦草造成重金属毒害。     

The influence of cyst nematodes and drought on potato growth. 3. Effects on carbon isotope fractionation

A good agreement was found between drought induced water-use efficiencies and carbon isotope fractionation of potato plants of the same age. Discrimination of¹³Carbon isotope increased with plant age and with the rank order of the distinguished plant organs. In young plants, nematode infection (contrary to drought) reduced¹³C discrimination but also reduced the water-use efficiency, leading to the conclusion that nematode infection, by slowing down the plant metabolism may have reduced the extrusion of¹³C. Towards senescence both nematode infection and drought reduced¹³C discrimination and increased water-use efficiencies indicating that then nematode caused water stress in the plants.     

家蚕品种茧层生产效率的调查及相关分析

251个家蚕品种的茧层生产效率平均为10．11％，最高12．19％，最低6．71％。二化性品种较高，一化性和多化性较低。中系二化种最高，平均10．54％，日二化种次之，平均10．39％，欧一化平均9.08％，日一化8．90％，多化平均8．72％。茧层生产效率与茧层量、茧层率、5龄一日茧层量成正相关，达显著或极显著水平；与5龄经过、全龄经过有正相关倾向，但不显著；与虫蛹率、死笼率、全茧量、消化率无一致倾向的相关。还调查了消化率和茧层转换率。     

非洲猪瘟病毒无标签重组B602L抗原制备与鉴定

为了获得无标签重组抗原用于非洲猪瘟病毒(ASFV)抗体检测,本研究将B602L与类弹性蛋白多肽(ELP)基因进行融合表达,利用简单、经济的相变循环(ITC)纯化ELP-B602L融合蛋白,用烟草蚀纹病毒(TEV)蛋白酶活性包涵体切除ELP标签,再用ITC回收重组B602L蛋白;用抗体阳性猪血清对重组B602L进行鉴定;以重组B602L蛋白为包被抗原进行ELISA鉴定。结果显示重组大肠杆菌能正确表达ELP-B602L融合蛋白,纯化融合蛋白纯度大于85%;TEV蛋白酶活性包涵体切除ELP标签的效率大于90%,回收的重组B602L蛋白纯度大于90%,能与抗体阳性猪血清反应;以重组B602L蛋白为包被抗原建立的ELISA与抗体阳性血清反应为阳性,与抗体阴性血清反应为阴性,OD450值与血清稀释倍数具有线性相关性。     

利用酵母双杂交膜系统筛选与小麦蓝矮植原体免疫膜蛋白互作的异沙叶蝉蛋白

正小麦蓝矮病(Wheat blue dwarf,WBD)是我国西北麦区一种重要植原体病害,在我国西部地区危害严重。该病害由异沙叶蝉(Psammotettix alienus L.)专化性传播,介体传毒成为病害流行的重要中心环节[1]。本实验室前期通过免疫荧光标记研究发现WBD植原体免疫膜蛋白(immunodominant membrane protein, IMP)与介体异沙叶蝉肌动蛋白互作,说明IMP在植原体传播和致病过程中起关键作用。     

Yield estimation of 1-year-old asparagus grown using rootstock-planting forcing culture

Rootstock-planting forcing culture was developed in asparagus to harvest spears even during the seasons when the plants become dormant, but the demand for them high. In this study, cumulative hours during which the air temperature remained lower than 5°C, i.e. chilling hours (CHs), were calculated to determine dormancy breakage for asparagus cultures. We also measured CIELab colour values for cut stems immediately before rootstock digging, and determined whether they could be substituted and/or compensated for CHs while evaluating asparagus plant productivity in different low-temperature backgrounds, and obtained regression equations for yield estimation. Asparagus seedlings were cultivated in seven different regions across Japan and brought to the study site for harvesting. Our regression equation based on CHs and rootstock weight for yield estimation had relatively high fitness (adjusted R² = 0.5795). The colour values of cut stalks at rootstock digging can also be used to evaluate their productivity. These values can be useful in regions where CHs cannot be determined, although their effectiveness was slightly lower than that of CHs of areas adjacent to the study sites.     

凤尾蕨科植物蜈蚣草的组织培养

[目的]对凤尾蕨科植物蜈蚣草的组织培养方法进行研究。[方法]以野外采集的叶芽为外植体,以1/2 MS＋3%蔗糖＋0.7%琼脂为基本培养基,通过激素配比试验,筛选蜈蚣草愈伤组织诱导和继代培养的培养基配方。[结果]从外植体诱导出愈伤组织最佳培养基配方为1/2 MS＋3%蔗糖＋0.7%琼脂＋0.5 g/L PVP＋0.1 mg/L KT＋0.5 mg/L 2,4-D;从愈伤组织诱导出GGB和从GGB诱导出幼苗最佳培养基配方为1/2MS＋3%蔗糖＋0.7%琼脂＋0.5 g/L PVP＋1.0 mg/L KT＋0.01 mg/L 2,4-D;另外,使用1/2 MS＋3%蔗糖＋0.7%琼脂＋0.5 g/L PVP＋0.5 mg/L 2,4-D做继代培养可以使愈伤组织持续增殖。[结论]研究直接使用野外材料组织培养,简化了试验步骤,是一种方便快速的蜈蚣草组培方法。