紫花苜蓿叶片对硒胁迫的生理响应研究

以紫花苜蓿"大叶苜蓿""维多利亚""阿尔冈金"为试材,采用土培法,研究了硒胁迫对3种紫花苜蓿叶片酶促防御系统的超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、过氧化物酶(POD)和抗坏血酸过氧化物酶(APX)活性及丙二醛(MDA)、可溶性蛋白质含量的影响。结果表明:在100μmol·L-1Se时,硒胁迫显著提高了3种紫花苜蓿叶片SOD、APX、POD、CAT活性和可溶性蛋白质含量。Se为900μmol·L-1时,SOD、APX活性和可溶性蛋白质、MDA含量显著增加,而CAT、POD活性显著降低。不同品种间抗氧化酶活性相比,"大叶苜蓿"抗氧化酶SOD、APX、POD、CAT活性显著高于其它2个苜蓿品种,且差异显著。综合评价表明,"大叶苜蓿"的抗氧化能力最强,其次为"阿尔冈金","维多利亚"苜蓿的抗氧化能力最差。     

红托鹅膏生态环境研究

以武定狮山为样点,对红托鹅膏的生境进行了初步研究。结果表明,红托鹅膏主要生长于滇青冈、滇石栎等阔叶林或针（云南松）阔混交林中,其生长的土表层疏松、透气、潮湿,具有厚4 cm~9 cm的腐叶层,有稀薄的阳光透入,土层有机质含量高,矿物质养分丰富,土壤容重较小,偏微酸性（pH5.4~5.7）。子实体分化发育较适宜的温度为19℃~21℃,空气湿度为75%~86%,土壤含水量为38%~50%,并每天保证1 300 lx~2 000 lx的光照2 h~5 h,200 lx~900 lx的弱光6 h~9 h。     

苜蓿冰结构蛋白抗冻活性及功能性研究

以"肇东"紫花苜蓿干草为原料,采用磷酸盐缓冲溶液法提取苜蓿冰结构蛋白,研究了温度、pH值、NaCl及蔗糖浓度不同提取因素对苜蓿冰结构蛋白功能的影响。结果表明:苜蓿冰结构蛋白溶液的冰晶体积小、数量多、形状不规则,具有很好的抗冻活性;对苜蓿冰洁构蛋白的功能性进行测定,随着溶液pH值的增加,其溶解指数、持水性、乳化性和乳化稳定性、起泡性与泡沫稳定性都呈现出逐渐增大的趋势;随着NaCl浓度的增加,其起泡性和泡沫稳定性先增加后降低、乳化性和乳化稳定性呈现下降趋势;随着蔗糖浓度的增加,其蛋白起泡性和泡沫稳定性逐渐增加,而乳化性和乳化稳定性影响不大。     

Relationship between Th1/Th2 balance and CD28/CTLA-4 expression on peripheral blood T cells in patients with systemic lupus erythematosus

AIM: To investigate the pattern of Th1/Th2 balance in systemic lupus erythematosus(SLE) patients and the relationship between CD28/CTLA-4(cytotoxic T-lymphocyte antigen-4) molecule expression and Th1/Th2 balance.METHODS: Eighteen SLE patients met the ARA 1997 updated SLE criteria were selected in the study. According to Bombardier's SLEDAI criteria, all patients were classified into two groups: active group(12 cases) and static group(6 cases). Fourteen normal individuals, matched for age and sex of the patients, served as controls. The peripheral blood mononuclear cells(PBMCs) were isolated by density gradient centrifugation and cultured in RPMI-1640 culture medium. After treated with PMA(5 μg/L) and ionomycin(500 μg/L) for 72 h, the PBMCs were collected, the contents of IFN-γ and IL-10 in the supernatant of cultured PBMCs were detected using enzyme linked immunosorbent assay(ELISA). The expression of CD28 and CTLA-4 molecules on T cells were detected by flow cytometric technique with double staining by FITC or PE labeled monoclonal antibodies. RESULTS: The level of IL-10 was higher in the PBMCs of active and static SLE patients(351.29 ng/L±153.31 ng/L and 319.37 ng/L±153.39 ng/L) than that in controls(254.48 ng/L±120.69 ng/L), but the difference did not reach statistical significance(P>0.05). The level of IFN-γ was significantly lower in the PBMCs of active SLE patients(25.76 ng/L±16.09 ng/L) than that in controls(50.71 ng/L±27.92 ng/L, P＜0.05). The ratio of IL-10/IFN-γ was significantly higher in active SLE patients(18.74±13.77) than that in controls(6.66±4.95, P<0.05). Either before or after culture, the expression of CD28 molecule on CD3⁺and CD8⁺ T cells from all SLE patients was not remarkably different from that in the cells of controls. Before culture, the expression of CTLA-4 molecule on CD3⁺T cells of active SLE patients(0.79%+0.37%) was significantly lower than that in the cells of controls(1.31%+0.61%, P<0.05). After culture, the expression of CTLA-4 molecule on CD3⁺ T cells of SLE patients was still lower than that in the cells of normal controls without statistical significance(P＞0.05).The expression level of CD28 molecule on CD3⁺ or CD8⁺ T cells in active SLE patients and controls was not correlated with the levels of IFN-γ and IL-10 in the supernatants(P＞0.05). The level of CTLA-4 molecule expression on CD3⁺ T cells of active SLE patients was positively correlated with IFN-γ level(r=0.681, P<0.05), while was negatively correlated with IL-10 levels(r=-0.624,P＜0.05) and the ratio of IL-10/IFN-γ(r=-0.738, P<0.01). The level of CTLA-4 molecule expression on CD3⁺ or CD8⁺ T cells of controls showed no correlation with IFN-γ levels, while showed negative correlations with IL-10 level(r=-0.587, P<0.05; r=-0.563, P<0.05, respectively).CONCLUSION: There is a bias in the differentiation of Th0 cells towards Th2 in SLE patients. CTLA-4 probably plays an important role in this mechanism through suppressing the signal transmitted by CD28.     

新疆野生樱桃李SSR体系的建立及应用

为建立适宜新疆野生樱桃李的SSR分子标记技术体系,通过对PCR反应程序、反应体系(DNA模板量、引物浓度、Mg2+浓度、dNTP浓度、Taq酶用量)以及退火温度进行了探索,建立了适宜野生樱桃李的SSR-PCR反应体系。结果表明:在25μL反应体系中,Mg2+1.0 mmol/L、引物0.5μmol/L、dNTPs 0.20 mmol/L、TaqDNA聚合酶1.0 U、模板DNA 30 ng、退火温度为60℃。SSR扩增程序:94℃预变性5 min,35个循环(94℃30 s,60℃1 min,72℃1 min),72℃延伸10 min,可筛选出稳定性好、多态性高的SSR引物。     

三种灌木的蒸腾耗水特性研究

利用Lcro+便携式光合仪,对青岛市3种主要灌木树种连翘、金银木、锦带花的蒸腾耗水规律进行了测定和研究。结果表明:在水分充足条件下连翘呈典型的双峰曲线,金银木和锦带花呈单峰曲线;金银木在1 d的各时间段耗水速率明显高于连翘和锦带花的耗水速率;3种灌木晴天的耗水速率明显大于阴天的耗水速率;此外,在春季,连翘对土壤含水量比金银木、锦带花敏感。试验的结果能够为植物的合理选择、水分的量化管理提供科学依据。     

我国甜菜抗逆性研究进展

从盐、水、温度、病害等方面综述了甜菜抗逆性研究进展,并对其研究前景进行了展望。     

龙眼体胚发生早期miR166初级体的克隆与表达分析

为了解龙眼miR166初级体基因Pri-miR166 S53结构特点及其前体和成熟体在龙眼体胚发生早期的表达模式,采用SMART~(TM) RACE试剂盒和PCR扩增技术克隆龙眼体胚早期miR166基因的初级体(Pri-miR166 S53),确认其转录起始位点,并预测其含有的潜在ORF;利用龙眼基因组数据库提取其启动子序列,预测其含有的顺式作用元件;用实时荧光定量PCR对龙眼体胚发生早期及不同激素处理下的胚性愈伤组织中miR166基因的前体(Pre-miR166 S53)和成熟体(miR166a.2)表达模式进行分析。结果表明:获得长317 bp的Pri-miR166 S53基因初级体序列,对其进行翻译,得到一条长13个氨基酸序列的miPEP(MLCFVDALFLIST)。利用生物信息学软件分析Pri-miR166 S53基因的启动子序列发现,除了具有TATA/CAAT-box外,还含有生长素、脱落酸、乙烯、水杨酸、茉莉酸甲酯及spl、HSE等特异作用元件。实时荧光定量PCR分析表明,在2,4-D调控的龙眼体胚发生早期过程中,从胚性松散型愈伤组织发育到球形胚的过程中,Pri-miR166 S53基因的前体pre-miR166 S53和成熟体miR166a.2都表现为下调趋势;而在无2,4-D调控的龙眼体胚发生早期中,pre-miR166 S53和miR166a.2表达模式不同。此外,pre-miR166S53随ABA和乙烯处理浓度升高呈下调表达,而对不同浓度2,4-D处理无应答;miR166a.2随2,4-D、ABA处理浓度升高呈下调表达,而在乙烯处理下呈上调表达。上述研究结果提示miR166前体和成熟体在对外源激素应答的模式上并不呈简单线性相关,可能存在多层次、多方位的调控。     

海南省火龙果溃疡病菌对吡唑醚菌酯敏感性分析

【目的】监测分析海南省火龙果溃疡病菌对吡唑醚菌酯的敏感基线及敏感性水平。【方法】采用菌丝生长速率法测定吡唑醚菌酯对海南省8个火龙果种植市(县)采集到的59株火龙果溃疡病菌的EC₅₀值,采用R语言进行数据分析。【结果】吡唑醚菌酯对供试菌株的EC₅₀范围为0.2540~2.2233μg·mL^-1,EC₅₀均值为0.9841μg·mL^-1;以58株菌株EC₅₀连续性正态分布时的均值0.9627μg·mL^-1作为海南省火龙果溃疡病菌对吡唑醚菌酯的敏感基线,海南省火龙果溃疡病菌均对吡唑醚菌酯表现为敏感;除乐东和白沙地区间敏感性差异显著外,其他地区间敏感性差异均不显著。【结论】海南省火龙果溃疡病菌对吡唑醚菌酯的敏感性差异较小,对吡唑醚菌酯仍较为敏感。     

丛枝菌根真菌在不同基质中对枳苗生长及氮磷吸收的影响

为筛选出适合枳砧木苗生长的菌根菌种类及基质组合,促进丛枝菌根(arbuscular mycorrhizal,AM)真菌在柑桔砧木容器苗生产中的高效应用,选取腐叶土、麦糠和菌渣为主要基质,分别与园土和河沙按体积比2∶1∶1配成混合栽培基质,以摩西球囊霉Glomus mosseae、地表球囊霉G.versiforme、地球囊霉G.geosporum和透光球囊霉G.diaphanum为供试菌种,采用容器育苗方法,研究了4种丛枝菌根真菌及其组合处理在3种基质下对枳苗生长及其氮、磷吸收的影响。结果表明,自然接种菌根侵染率不超过40%,人工接种侵染率可高达80%;枳苗以地表球囊霉接种和腐叶土或麦糠组合的侵染率最高,分别为80.7%和78.7%;以地表球囊霉接种和麦糠组合促进生长的效果最显著,菌根依赖性达到342.4%,生物量比对照高出278%,该组合促进植株氮、磷吸收的效果也最显著,其吸氮总量比对照高482.0%,吸磷总量比对照高553.8%。适合枳容器苗生长的最佳AM真菌和基质组合是地表球囊霉+麦糠混合基质。