日本百脉根LjbHLH34基因克隆及耐旱功能鉴定

干旱是影响植物生长发育的重要环境因素。本研究分析了日本百脉根抗旱相关基因LjbHLH34的耐旱功能，初步解析其响应干旱胁迫的分子机制，以期为百脉根抗旱分子育种提供理论基础。本研究克隆得到的LjbHLH34基因大小为711 bp、编码236个氨基酸，属bHLH转录因子家族成员。系统进化树分析显示，LjbHLH34蛋白与拟南芥bHLHⅣ亚家族中AtbHLH34和AtbHLH104亲缘关系较近。实时荧光定量分析表明LjbHLH34在日本百脉根的根中表达量最高，叶中次之，茎中最少，暗示其在日本百脉根多个组织中发挥作用；同时LjbHLH34基因也受聚乙二醇(PEG)和脱落酸(ABA)诱导表达。在酵母中检测发现LjbHLH34具有转录激活活性；亚细胞定位试验表明LjbHLH34蛋白定位于细胞核中。将LjbHLH34基因转入拟南芥获得过表达株系。在200 mmol·L^-1甘露醇胁迫下，LjbHLH34转基因拟南芥的根长明显长于野生型。干旱处理后，野生型拟南芥比转基因拟南芥萎蔫程度更加明显，而转基因株系的相对含水量和超氧化物歧化酶(SOD)活性显著高于野生型，丙二醛(MDA)积累...     

水稻颖花持续开放突变体sostenuto floret opening(sfo1)的鉴定与基因定位

水稻颖花开放是其生殖发育一个关键生理过程,对受精和随后种子发育具有重要影响。本文报道了一个与水稻颖花开放相关的突变体,来源于籼稻保持系西农1B的EMS(ethyl methane sulfonate)诱变群体。该突变体表现为开颖后浆片失水萎缩过程缓慢,内外稃持续开裂不闭合,暂命名为水稻颖花持续开放sostenuto floret opening 1(sfo1)突变体。遗传分析表明sfo1性状受1对隐性单基因控制,利用群体分离分析法(bulked segregation analysis,BSA)将SFO1基因定位在第5染色体SSR标记RM1054和IN/DEL标记ZTQ51之间,物理距离113 kb,含注释基因15个。本研究结果为SFO1基因的图位克隆和功能研究奠定了基础。     

Effect of tetrandrine on radiosensitivity of nasopharyngeal carcinoma cells

AIM: To investigate the mechanism of the radiosensitizing effect of maximum non-cytotoxic doses of tetrandrine (Tet) on nasopharyngeal carcinoma cell lines CNE1 and CNE2.METHODS: The cells were treated with ma-ximum non-cytotoxic doses of Tet (for CNE1 cells at 1.5 μmol/L and for CNE2 cells at 1.8 μmol/L), irradiation at 4 Gy, or combination of irradiation and maximum non-cytotoxic doses of Tet. The cell cycle distribution was analyzed by flow cytometry. The protein levels of γ-H2AX, cleaved caspase-3, p-CDC25C, CDK1, p-CDK1, cyclin B1, ERK and p-ERK were determined by Western blot.RESULTS: The expression of γ-H2AX was increased in CNE1 cells and CNE2 cells after combined treatment with irradiation and maximum non-cytotoxic doses of Tet. The percentages of CNE1 cells and CNE2 cells at G₂/M phase in irradiation group were (18.09±0.42)% and (18.48±1.32)%, respectively, which were decreased to (15.88±1.04)% and (13.80±0.82)% in combined treatment group, respectively (P<0.05). Combined treatment enhanced the increase in the protein level of cleaved caspase-3 caused by irradiation. The protein levels of p-CDC25C and p-CDK1 were increased in a dose-dependent manner by Tet treatment (P<0.05), while the expression of CDK1 showed no difference among different doses of Tet treatments. The protein levels of p-CDC25C, p-CDK1 and CDK1 showed no difference after the treatment with maximum non-cytotoxic doses of Tet. The combined treatment with irradiation and the maximum non-cytotoxic doses of Tet decreased the protein levels of p-CDC25C and p-CDK1 (P<0.05), increased the expression of cyclin B1, and had no influence on the expression of CDK1 (P<0.05). The combined treatment resulted in an increase in the protein level of p-ERK1 (P<0.05).CONCLUSION: The maximum non-cytotoxic doses of Tet enhance the DNA damage and apoptosis in CNE1 cells and CNE2 cells caused by irradiation, and the mechanism might be associated with ending of G₂/M arrest via activation of ERK/CDC25C/CDK1/cyclin B1 pathways.     

假禾谷镰孢侵染小麦后3种植物激素相关基因的差异表达分析

假禾谷镰孢(Fusarium pseudograminearum)是我国近年新发现的小麦病原真菌,本研究目的是揭示植物激素信号传导途径中相关基因表达对假禾谷镰孢侵染的响应。利用假禾谷镰孢野生菌株WZ2-8A侵染小麦品种周麦24,对接种后5 d和15 d样品进行转录组测序,分析差异表达基因,并对候选基因进行q RT-PCR验证。假禾谷镰孢侵染后,小麦幼苗生长受到明显抑制,根长、株高、根重和地上部分鲜重均显著降低。转录组分析结果表明,植物激素信号传导途径中有29个基因差异表达,涉及到生长素、细胞分裂素和脱落酸3种植物激素。在接种后5 d有11个差异表达基因,其中2个上调,9个下调;在接种后15 d共有25个差异表达基因,其中8个上调,17个下调。在生长素信号传导途径中,生长素输入载体AUX1差异表达,影响生长素的极性运输,从而影响小麦根部的细胞伸长。在细胞分裂素信号传导途径中,起正调控作用的B-ARR上调表达,推测其促进细胞分裂素的信号传导,从而抑制细胞分裂,与生长素协同作用,造成小麦的长势减弱。在脱落酸途径中,脱落酸受体PYR/PYL下调表达;起到负调控作用的PP2C相关基因均上调表达。脱落酸使植物对真菌和细菌的抗性起到负调控作用,其信号传导途径与茉莉酸/乙烯途径相互拮抗,脱落酸信号传导途径的阻遏可能会使茉莉酸/乙烯途径信号通路打开。q RT-PCR结果基本能够和转录组测序结果相拟合,说明在假禾谷镰孢侵染胁迫下,脱落酸的信号传导被抑制可能是中抗品种周麦24对假禾谷镰孢产生一定的抗性的生理基础。     

Chronic hypoxia enhances aggressiveness of MCF-7 breast cancer cells through EMT

AIM: To investigate the effects of chronic hypoxia on the aggressiveness of MCF-7, a human breast cancer cell line, and the underlying mechanisms.METHODS: MCF-7 cells were cultured under hypoxia (1% O₂, 5% CO₂ and 94% N₂) or control (95% O₂ and 5% CO₂) condition. The viability, proliferation, and invasion and migration abilities of the MCF-7 cells were determined by MTT assay, CCK-8 assay, cell counting, and cell invasion and migration assays. Anchorage-independent growth and the alteration of cellular polarization of the MCF-7 cells were tested by soft agar colony formation assay and Matrigel-3D culture assay, respectively. The effects of chronic hypoxia on the growth and metastasis of MCF-7 cells in vivo were investigated by xenograft in nude mice. The morphological changes of the MCF-7 cells were observed under an inverted microscope. Hypoxia-induced alterations in the levels of hypoxia inducible factor-1 (HIF-1) and phosphorylated glycogen synthase kinase-3β (p-GSK-3β) as well as epithelial-mesenchymal transition (EMT) molecules, such as E-cadherin, N-cadherin, vimentin, matrix metalloproteinase (MMP)-3 and MMP-9, were determined by Western blot.RESULTS: Chronic hypoxia significantly increased the viability, proliferation, and invasion and migration abilities of MCF-7 cells in vitro, enhanced the anchorage-independent growth, facilitated cellular polarization alteration in Matrigel-3D culture, and promoted cancer metastasis in vivo. Hypoxia up-regulated HIF-1, activated GSK-3β, down-regulated E-cadherin and increased the protein levels of N-cadherin, vimentin, MMP-3 and MMP-9. CONCLUSION: Chronic hypoxia enhances the aggressiveness of breast cancer cells probably through EMT.     

茎瘤芥叶栽培金针菇试验

以茎瘤芥叶、棉籽壳为主要原料栽培金针菇,筛选最佳栽培配方,拟为茎瘤芥叶的循环利用提供参考依据。结果表明:金针菇最适栽培配方的质量百分数为24%茎瘤芥叶、60%棉籽壳、10%麸皮、5%玉米粉、1%石膏;以此配方栽培金针菇,每袋产量最高为248.48g、生物学效率为88.74%,投入产出比为1∶3.30,比全棉籽壳栽培金针菇降低成本21.43%,提高毛利润16.44%。     

基于溯源技术的蔬菜基地管理系统开发与应用

蔬菜是我国重要的农产品,其质量安全问题关系着国计民生,一直是社会关注的重点。当前我国的蔬菜产品质量安全问题仍然十分广泛和严重,问题蔬菜的来源去向不明、责任不清,企业对蔬菜种植基地的管理缺乏有效的信息管理手段,消费者对购买蔬菜缺乏有效可靠的信息获取途径。因此,从信息技术的角度构建了一个基于溯源技术的蔬菜基地管理系统,详细介绍了蔬菜溯源码的设计、系统的开发环境、业务流程与主要功能,并重点阐述以规程控制为农业生产指导的蔬菜安全管理模式。系统的开发与应用,实现了"问题蔬菜"的企业内部溯源,以及消费者通过手机查询蔬菜溯源信息,为蔬菜企业提供了对基地种植进行质量管理的信息化工具,为政府提供了一种溯源系统解决方案。     

大学生学习压力的调查分析与对策——以云南农业大学为例

随着我国高等教育的发展,大学教育已从精英教育向大众化教育转变,但学习依然是大学生的主要任务,学习压力伴随着大学生,特别是面对严峻的就业形势,大学生学习压力有增无减。本文通过对云南农业大学大学生学习压力的实证调查,从大学生所承受的学习压力程度来了解大学生学习压力现状;从大学生学习压力来源中分析其产生学习压力的各种原因,并针对原因提出自我调节、高校积极开展大学生心理健康教育活动、家长对大学生树立合理期望值、社会为大学生创造良好外部环境的"四位一体"调控对策,缓解大学生学习压力。     

陆地棉SRO基因家族的鉴定及表达分析

SRO是植物特有的类RCD-ONE蛋白家族,在植物抵御各种生物和非生物胁迫过程中具有重要的作用。本研究利用生物信息学方法从陆地棉遗传标准系TM-1(Gossypium hirsutum L.acc.TM-1)基因组中鉴定到12个SRO基因家族成员。多重序列比对及进化树分析表明,12个陆地棉SRO均含有PARP和RST结构域,聚为3类,即A、B和C亚家族。转录组数据分析表明,TM-1全基因组中的Gh_D12G1442在茎中优势表达,Gh_D05G2064在萼片和雄蕊中优势表达;Gh_A05G3788在雄蕊中优势表达,Gh_A12G1318在雄蕊和纤维中优势表达;Gh_A12G2480、Gh_D12G2608、Gh_A05G2257和Gh_D05G2516在纤维中优势表达;Gh_A08G1390、Gh_D08G1685、Gh_A12G2663和Gh_D12G2054具有较为广泛的组织表达。逆境胁迫处理试验表明,Gh_A08G1390明显受冷处理和盐处理的诱导,Gh_D12G2054、Gh_D08G1685、Gh_A12G2663基因在不同的胁迫处理下受到不同程度的诱导。     

绿豆品种资源萌发期耐碱性鉴定

采用人工气候箱内培养皿培养,以混合碱Na HCO3∶Na2CO3(摩尔比)为9∶1模拟典型东北地区碱胁迫环境,在萌发期以50 mmol L–1溶液处理34份绿豆品种资源,蒸馏水处理为对照,每培养皿放30粒种子。培养第3天测定发芽势,第7天测定发芽率、下胚轴长、胚根长、下胚轴干重、胚根干重等指标,通过隶属函数法和聚类分析对参试材料耐碱性综合评价,并进行因子分析。利用隶属函数法对参试材料耐碱性排序表明,不同绿豆品种资源间表现出较大差异,聚类分析把参试材料按耐碱性强弱分为4大类,白绿11等9份材料为耐碱类型,公绿1号等19份材料为耐碱中间类型,吉绿3号等5份材料为碱敏感类型,潍绿7号为碱极敏感类型。因子分析结果表明,萌发指数、下胚轴干重、胚根长分别在萌发因子、生物量累积因子和伸长因子中的负荷量最大,可作为绿豆萌发期耐碱性鉴定的适宜指标。