梨灰霉病拮抗放线菌L-30的筛选、鉴定及作用机制研究

采用平板对峙培养法从果树根际土壤中筛选出对梨灰霉病菌(Botrytis cinerea)及10种常见果实采后病害具有抑制活性的放线菌株L-30,同时利用分子方法结合形态观察对其进行鉴定,并测定其对B.cinerea孢子萌发的抑制作用、对‘皇冠’梨的诱导抗性和果实防御酶系活性的影响。结果表明,菌株L-30对梨灰霉病菌的活体抑制活性达到40%以上,基于16S rDNA核酸一致性和系统发育分析及培养特性证明L-30为链霉菌属(Streptomyces)的多产色链霉菌(S.polychromogenes)。L-30稀释100倍发酵液对梨灰霉病孢子萌发抑制率为52.04%。L-30处理皇冠梨果实能显著降低梨灰霉病菌的扩展,并提高果实的过氧化物酶(POD)、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)和苯丙氨酸解氨酶(PAL)活性。     

高效实用型黄瓜嫁接技术——生长点斜插法

为了提升黄瓜嫁接苗对设施栽培连作障碍的耐受能力,提高幼苗的抗病性与抗逆性,同时确保嫁接成活率,增加产品产量及品质,在传统的黄瓜嫁接方法基础上,改进出高效实用的生长点斜插嫁接法,为此创新出了专用嫁接工具——β形铝合金嫁接签,并申报了实用新型专利（专利号： 201811595026.X）;为了提高嫁接苗的成活率,在对砧穗进行特定处理的同时,还探索出了嫁接苗生长最佳环境的保障装置——育苗钢架。通过育苗基质的营养控制、省略嫁接夹具的插接方法及保温保湿的育苗钢架等系列配套技术的使用,使得生长点斜插嫁接技术具备了突出的实用性与高效性,十分适合在实际生产中推广使用。     

不同土壤深度棉花根系形态变化规律及与矿质元素的关系

【目的】研究不同土壤深度下棉花根系形态变化规律,分析与矿质元素的响应关系,及矿质元素对根系形态的调控作用,为构建合理棉花根系构型提供理论依据。【方法】测试37份棉花品种的根系长度、根系表面积、根系体积、根系生物量、根系氮、磷、钾元素,根据公式计算比根长、比根面积、比根体积、根系密度,研究矿质元素对根系形态的调控关系。【结果】位于0~10 cm土壤中,由于氮素作用,促使比根长、比根面积、比根体积增加,轻微抑制根系密度生长。在10~20 cm土壤中,氮素促使比根长、比根面积增加,抑制根系密度的生长及比根体积的增加。在20~30 cm土壤中,氮素对比根长、比根面积、根系密度的作用效果不明显,而对比根体积表现为负相关关系。在30~50 cm土壤中,氮素对比根长、比根面积、比根体积、根系密度表现为正相关关系。在50~60 cm的土壤中,比根长、比根面积、比根体积、根系密度表现为负相关关系。第一主成分与氮素、比根长、比根面积、比根体积的相关性较高,第二主成分主要与根系密度的相关性较高,第三主成分主要与磷、钾元素的相关性较高。磷、钾元素与第一主成分的相关性较高,氮素与第三主成分的相关性较高。【结论】根系在不同土壤深度下,比根长、比根面积、比根体积逐渐增大,单位质量根系含氮量不断减少。根系形态性状主要受氮素调控,磷、钾元素对根系形态性状的调控作用较小。     

山羊和绵羊嗜血支原体PCR检测方法的建立与应用

为建立羊嗜血支原体(M.ovis)病快速检测方法,本实验根据GenBank中登录的羊M.ovis 16S rRNA基因序列设计一对引物,以山羊和绵羊M.ovis基因组DNA为模板,建立M.ovis PCR检测方法,并进行特异性、敏感性及临床应用试验。结果显示,建立的M.ovis PCR检测方法扩增片段大小为508 bp,与GenBank中M.ovis参考株同源性达99%以上,该方法与猪嗜血支原体、牛嗜血支原体等病原体无交叉反应,最低检测40个拷贝的DNA;通过对延边地区60份山羊和绵羊血液样本的检测结果表明,建立的PCR检测方法具有特异、敏感、准确等优点,完全适用于M.ovis的检测。     

马疱疹病毒1型主要毒力基因分析及TK基因缺失载体的构建

为了解新疆马疱疹病毒1型（EHV-1）主要毒力基因遗传进化情况并构建TK基因缺失株,本研究以EHV-1 XJ2015株DNA为模板,对其主要毒力基因TK、gI和gE全长进行克隆、测序及生物信息学分析,并扩增TK基因左右重组臂TKL和TKR,构建质粒pUC-TKLR,将扩增后的增强绿色荧光蛋白（EGFP,含有CMV+polyA）插入pUC-TKLR质粒,构建TK基因缺失打靶质粒。TK、gI和gE基因同源性分析结果显示,XJ2015株与国外EHV-1分离株TK、gI和gE基因同源性均较高,分别为99.8%~100.0%、99.6%~100.0%和99.9%~100.0%;与EHV-3分离株同源性均最低,分别为72.9%、59.4%和62.1%;遗传进化分析显示,3个基因均与国外EHV-1同属于一个遗传进化分支,与EHV-9和EHV-4进化关系较近,但与EHV-3进化关系较远,表明XJ2015毒株与国外EHV-1毒株TK、gI、gE基因核苷酸上差异不明显,没有明显的地域性特征,功能基因保守且进化缓慢,同源基因功能相同或相近;经PCR扩增、酶切、测序及转染鉴定,本试验成功构建了用于TK基因缺失的打靶质粒pUC-TKLR-EGFP。通过对EHV-1主要毒力基因的分析及TK基因缺失打靶载体的构建,为新疆地区马鼻肺炎流行病学调查分析、TK基因缺失株的构建提供理论依据。     

Effects of different active constituents of Gynostemma pentaphyllum on mitochondrial energy metabolism-related proteins in endothelial cells induced by ox-LDL

AIM To investigate the effects of different components of Gynostemma pentaphyllum ［gypenosides (Gps), gypenoside XLIX (GpXLIX) and ginsenoside Rb3 (GRb3)］ on mitochondrial energy metabolism-related proteins in endothelial cells induced by oxidized low-density lipoprotein (ox-LDL). METHODS EA.hy926 cells were divided into control group, model group, Gps group, GpXLIX group and GRb3 group. The cells in control group were cultured only in DMEM complete medium. The cells in model group were treated with 100 mg/L ox-LDL for 48 h. The cells in Gps group, GpXLIX group and GRb3 group were treated with 100 mg/L ox-LDL for 24 h, and then treated with Gps, GpXLIX and GRb3 at 100 mg/L for another 24 h, respectively. The ATP content in each group was detected by ELISA. The expression levels of mitochondrial energy metabolism-related proteins, cytochrome C oxidase subunit 5a （Cox5a）, NADH:ubiquinone oxidoreductase core subunit S1 （Ndufs1）, ATP synthase F1 subunit alpha (ATP5a) and cytochrome C (Cyt C）, were determined by Wes automatic Western blot quantitative analysis system and Western blot. RESULTS Compared with control group, the ATP content in model group was decreased (P<0.01). After drug intervention, the ATP content increased to different degrees in Gps group, GpXLIX group and GRb3 group (P<0.01). The results of Wes automatic Western blot quantitative analysis system were consistent with those of Western blot. These results showed that compared with control group, the protein expression of Cox5a, Ndufs1 and ATP5a in model group was decreased, and the protein expression of Cyt C was increased (P<0.01). After intervention, the protein expression of Cox5a, Ndufs1 and ATP5a was increased and the protein expression of Cyt C was decreased in Gps group, GpXLIX group and GRb3 group (P<0.05 or P<0.01). Among them, the effect of Gps on the protein expression of Cox5a, Ndufs1 and Cyt C was significantly stronger than those of the 2 monomer components, and the effect of GRb3 was found to be superior in the 2 monomer components. The effect of GpXLIX on ATP5a protein was superior to the other 2 components. CONCLUSION Gynostemma total saponins and related active ingredients protect ox-LDL-induced endothelial cells by affecting mitochondrial energy metabolism-related proteins, thereby preventing and treating atherosclerosis.     

小麦条锈菌DNA提取方法比较及RAPD反应体系优化

分别采用CRAB法和CTAB/SDS法进行小麦条锈菌gDNA的提取,并对两种方法进行比较分析.结果表明:CTAB法具有较高的得率,其优化RAPD反应体系为10×Reaction Buffer 2.5 μL,MgCl<,2>(25 mmol/L)2 μL,dNTP(2.5 mmol/L each)1.5μL,模板DNA(...     

NaHS引发提高裸燕麦种子活力的生理机制

硫化氢(H2S)信号在作物种子萌发中发挥着重要作用.为探讨外源H2S供体NaHS引发提高作物种子活力的作用及其生理机制,以裸燕麦种子为材料,分别用不同浓度NaHS(0、50、100、200、400、800、1600μmol·L-1)引发18 h和800μmol·L-1 NaHS引发不同时间(6、9、12、15、18、2...     

不同干燥方法对苜蓿干草调制的影响

针对影响人工干燥苜蓿的水分、密度、摆放方式和人工辅助干燥方式4个因素进行人工鼓风干燥处理,设计L9(34)正交试验,监测高水分苜蓿草捆的湿度变化情况,比较不同处理营养物质、相对饲用价值和经济指标.对比分析结果可知:自然晾晒苜蓿干草时,营养物质损失严重;苜蓿高水分打捆后人工继续鼓风干燥处理有利于保存苜蓿干草的营养价值,其中水分为30％～35％、密度为120kg/m3、间隔20cm摆放、两台鼓风机每日鼓风干燥3h为最优组合,所得苜蓿草捆的品质较好.     

Forest recovery after clear-cutting in Chinese pine(Pinus tabuliformis) plantations of North China

In Hebei Province of North China, forest was recovered with natural recruitment in plantations with large area of clear-cutting Chinese pine(Pinus tabuliformis). This study was aimed to demonstrate the dynamic characteristics of recruits during the natural recruitment. Both plot survey and the spatial point-pattern analysis were performed. Five developmental stages of natural recruitment were selected and studied, including 1 year before and 2, 5, 8, and 11 years after clear-cutting. Different slope aspects were also included. Natural recruitment was always dominated by Chinese pine with a proportion of higher than 90%. For plots of 1 year before clear-cutting on east-and north-facing slopes, recruit densities were 7886 and 5036 stems/hm2, the average heights were 0.78(±0.85) and 1.06(±1.15) m, and the average diameters at breast height(DBH) were 3.21(±1.38) and 2.91(±1.38) cm, respectively. After clear-cutting, recruit density was initially increased, then it was gradually declined with time; however, the variation of average DBH was contrary to that of recruit density. Both of them were no longer varied between 8 and 11 years after clear-cutting. The average height of recruits continued to increase after clear-cutting. For the plots of 11 years after clear-cutting on east-and north-facing slopes, average heights of recruits reached 2.00(±1.14) and 2.24(±1.20) m, respectively. The statuses of recruits on north-facing slopes were better than those on east-facing slopes after clear-cutting. Meanwhile, recruits on east-facing slopes were always aggregated at small scales, while spatial pattern of recruits varied with time on north-facing slopes. Moreover, forest was recovered more quickly by natural recruitment than by artificial afforestation after clear-cutting. The structural diversity was higher in naturally regenerated forests than in plantations of the same age. Our results demonstrated that clear-cutting of Chinese pine plantations recovered by natural recruitment has the potential to be an effective approach for establishing multifunctional forest.