花生种子特异启动子AHSSP1的克隆及功能分析

为丰富花生种子特异启动子资源,本研究利用PCR技术在花生基因组中克隆了种子贮藏蛋白基因PSC32的启动子AHSSP1,利用半定量RT-PCR检测了PSC32基因表达模式,借助NewPLACE在线分析了AHSSP1序列中存在的顺式作用元件,并构建了AHSSP1驱动GUS报告基因的表达载体,经农杆菌转化获得转基因拟南芥,经GUS组织化学染色鉴定了该启动子的功能。结果表明,PSC32基因957 bp长的启动子AHSSP1序列具备种子特异表达启动子特有的3个RY REPEAT元件。半定量RT-PCR分析发现,PSC32基因在花生成熟种子中表达,而在饱果成熟期根、茎、叶片、花、入土前的果针、成熟种子的果壳中均不表达。GUS组织化学染色发现,转基因拟南芥成熟种子以及萌发种子的子叶、下胚轴和胚根均能够被染上蓝色;长出真叶后,子叶和下胚轴仍能被染色,而根和真叶不能被染上蓝色;成年期转基因拟南芥的叶片也不能被染上蓝色。而野生型拟南芥整个生长时期均不能被染上蓝色。以上现象说明AHSSP1是一个种子特异启动子。本研究丰富了花生种子特异启动子的资源,对花生籽仁品质改良或以花生籽仁作为“生物反应器”的研究具有重要的应用价值。     

硅钙钾镁肥配施黄腐酸钾对土壤酶活性及桃幼树生长的影响

为探究硅钙钾镁肥配施黄腐酸钾对桃树生长的效应,以盆栽一年生瑞蟠21号毛桃(Amygdalus persica Linn.)为试验材料,设置3种不同施肥方式(T1:硅钙钾镁;T2:硅钙钾镁+黄腐酸钾;CK:空白对照),在桃幼树生长季进行追肥处理,研究硅钙钾镁肥及硅钙钾镁肥配施黄腐酸钾对土壤酶活性、土壤养分状况及桃幼树根系构型和植株生长的影响。结果表明,与CK相比,T1、T2均显著提高了桃幼树生长季各时期土壤酸性磷酸酶、脲酶和过氧化氢酶活性,以及土壤中碱解氮、有效磷、速效钾和有效硅等含量,且配施黄腐酸钾后作用更明显。与CK相比,T1、T2细根比重分别提高了11.2%和26.2%,根系活力分别提高了12.2%和18.1%,根系总长度分别增加了28.1%和61.9%,且有效延缓了根系衰老;T1、T2植株总干重分别提高了1.2倍和2.0倍,根冠比分别提高了9.4%和17.0%,且差异显著。与CK相比,T1、T2各器官中全氮、全磷、全钾含量均显著增加,地上部硅含量分别提高了61.3%和91.9%,地下部硅含量分别提高了62.3%和102.5%,且差异显著。同时,T1、T2桃幼树叶片叶绿素含量和净光合速率均显著提高。综上所述,硅钙钾镁肥可有效提高土壤酶活,促进桃幼树细根的生长,延缓根系衰老进程,增加氮、磷、钾、硅等营养元素积累量,提高叶片光合速率,促进植株生长;硅钙钾镁肥配施黄腐酸钾后,提高了土壤有效硅及植株硅含量,比单施硅钙钾镁肥效果更加显著。本研究结果为桃树合理施用硅钙钾镁肥提供了理论依据。     

60Co-γ射线和电子束辐照灭菌对葛根提取物抗氧化活性及指纹图谱的影响

为评价⁶⁰Co-γ射线和电子束辐照灭菌对葛根提取物品质的影响效应及其差异性,采用不同剂量的⁶⁰Co-γ射线(0、2.8、5.7、8.3、10.6 kGy)和电子束(0、2.4、5.1、7.7、10.3 kGy)辐照处理葛根提取物,研究辐照对其微生物总数、抗氧化活性及指纹图谱的影响。结果表明,⁶⁰Co-γ射线和电子束辐照均能有效降低葛根提取物中需氧菌总数、霉菌和酵母菌总数,随着吸收剂量增大,抑制作用增强。⁶⁰Co-γ射线和电子束辐照吸收剂量分别为5.7 kGy和5.1 kGy时,葛根提取物均达到《中国药典》所规定的微生物限度标准(需氧菌总数为10³ CFU·g^-1、霉菌和酵母菌总数为10² CFU·g^-1);吸收剂量分别为5.7 kGy和7.7 kGy时,需氧菌总数、霉菌和酵母菌总数均降至10 CFU·g^-1以下。此外,大肠埃希氏菌(Escherichia coli)与沙门氏菌(Salmonella)均未检出。2种辐照方式均对葛根提取物的DPPH自由基(DPPH·)清除率、羟自由基(·OH)清除率以及总还原能力无显著影响(P>0.05)。不同剂量⁶⁰Co-γ射线和电子束辐照处理葛根提取物特征峰的相似度达到1.0,且相对保留时间的相对标准差(RSD)均小于0.5%,表明2种辐照方式对葛根提取物指纹图谱均无显著影响(P>0.05)。综上所述,⁶⁰Co-γ射线和电子束辐照对葛根提取物微生物具有较强的抑制作用,2种辐照方式均不会对葛根提取物的抗氧化活性及指纹图谱产生显著影响,不影响葛根提取物的有效性及质量稳定性。该研究为2种辐照技术,特别是电子束辐照在葛根药材、饮片及其中成药制剂灭菌中的合理应用提供了科学参考。     

海南粗榧内生真菌F127发酵液的次级代谢产物研究

对海南粗榧(Cephalotaxus hainanensis Li.)内生真菌F127发酵液化学组分进行研究。用多种色谱技术对F127代谢产物进行分离纯化,以LC-MS、ESI-MS和超导核磁共振分析鉴定化合物结构,最后采用MTT法测定化合物的抗肿瘤活性。从海南粗榧内生真菌F127的发酵液中分离得到5个单体化合物,分别鉴定为oblongolide T(1)、sorbicillin(2)、邻苯二甲酸二丁酯(3)、phomopsolide B(4)、6,8-二羟基-3-甲基-3,4-二氢异香豆素(5),且化合物1、4、5对肿瘤细胞K562、NB4、HL-60、Hep G-2和Lovo表现出不同抑制活性。化合物4对5株肿瘤细胞均表现出抑制活性,对K562、NB4、HL-60抑制效果显著,IC50值分别为3.35、0.014和0.16μg/m L,对Hep G-2和Lo Vo只有温和抑制作用;化合物1对K562、NB4、HL-60抑制作用良好,IC50值分别为51.82、54.25和29.31μg/m L;化合物5对NB4和Hep G-2则具有一定抑制活性;化合物2和3对测试细胞株未表现出抑制活性。5个化合物均是首次从海南粗榧内生真菌中分离得到,并首次报道了化合物1、4对K562、NB4、HL-60的优良细胞毒活性,为进一步研究海南粗榧内生真菌中的活性天然产物奠定了基础。     

Characteristics of starch synthesis and grain filling of common buckwheat

The common buckwheat ‘cv. Fengtian 1’ (FT1), ‘cv. Yuqiao 4’ (YQ4), ‘cv. Dingtian 2 (DT2)’, and ‘cv. Tongliao (TL)’ were selected to investigate the characteristics of starch synthesis and grain filling. The chlorophyll and chlorophyll a/b contents of leaves in the third stem showed a steady downward trend with the growth of common buckwheat. Whereas, FT1, YQ4 and DT2 showed higher chlorophyll content than that of TL at 7d and 14d. The activity of soluble starch synthase(SSS) and adenosine diphosphate glucose pyrophosphorylase (ADPGP) in the four tested varieties showed a rapid upward trend until 14 days after heading but then rapidly decreased until 21 days after heading. The average adenosine diphosphate glucose pyrophosphorylase activities of FT1, YQ4 and DT2 were higher than TL, while the average soluble starch synthase activities were similar among the four tested varieties. Consistently, the four tested varieties showed rapid accumulation of starch, amylose, and amylopectin during the prefilling stage, but no difference of starch content was found at maturity. The filling process of the four common buckwheat varieties exhibited an “S” curve. The Richards equation was utilized to evaluate the grain-filling process of common buckwheat. YQ4 showed the largest values of initial growth power (R₀). The time reaching the maximum grain-filling rate (t_max.G) of FT1 was the longest, and the maximum grain-filling rate (G_max) of DT2 was the fastest.     

果蔬籽油生物活性研究进展

果蔬籽油是由果蔬籽经压榨或萃取工艺制得的植物油脂。果蔬籽油含有丰富的维生素、多不饱和脂肪酸、有机酸等活性成分,这些成分赋予了果蔬籽油抗氧化、抗癌、调节血脂和调节免疫系统等生物活性。本文综述了果蔬籽油生物活性的研究进展,旨在为果蔬籽油的综合开发和利用提供理论依据和指导。     

HXKs基因在香蕉果实后熟过程中功能初探

香蕉是世界主要水果之一,对乙烯非常敏感,属于呼吸跃变型果实。目前,香蕉果实后熟机理还未完全探明,后熟过程中己糖激酶(HXK)与乙烯的关系还不清楚。通过BLAST比对从香蕉基因组数据库中发现HXK基因家族的11个成员,经克隆获得这些基因的序列,并研究HXKs基因在香蕉果实自然后熟过程中的转录表达谱,重点研究乙烯利、甘露糖、NAG和1-MCP对HXKs基因表达、HXK酶活和内源乙烯生物合成的影响。结果表明:在香蕉果实后熟过程中HXKs基因呈现差异性表达,乙烯利上调大多数HXKs基因的表达和HXK酶活,1-MCP的作用正好相反,这表明外源乙烯正作用于HXK。另一方面,甘露糖加快内源乙烯生物合成,NAG却推迟内源乙烯生物合成,这说明HXK正作用于内源乙烯生物合成。所以,在香蕉果实后熟过程中乙烯和HXK之间存在相互促进的关系。这将为进一步阐明香蕉果实后熟机制提供理论依据,也将为香蕉果实保鲜新技术的挖掘提供新思路。     

Resistance monitoring and enzyme activity in three field populations of cowpea aphid (Aphis craccivora) from Egypt

The cowpea aphid (Aphis craccivora) is one of the most important sucking insect pests attacking certain legumes in Egypt particularly faba bean, cowpea and pea. In this study we monitored the resistance level of three field populations of A. craccivora to seven insecticides belonging to three different chemical classes (organophosphates, carbamates and neonicotinoids). The three populations were collected from three governorates in Egypt namely Dakahlia, Qalyobia and Beni Suef. Diagnostic concentrations (LC₉₀ values for susceptible strain) for each insecticide were established using a leaf dipping technique. Resistance monitoring showed that the field population from Dakahlia was highly resistant to all the tested insecticides. In a similar manner, the population from Qalyobia was also resistant to all insecticides except for fenitrothion to which only moderate resistance was observed. The field population from Beni Suef exhibited a lower level of resistance to all the seven tested insecticides.Biochemical assay showed that esterase activity in these three field populations was generally higher as the enzyme activity ratio ranged from 4.3 to 7.8 fold more than that for the susceptible strain. The activity of the other measured detoxifying enzymes (glutathione -S- transferase and mixed function oxidases) was moderate in the populations from Qalyobia and Dakahlia. Nevertheless, the enzyme activity in A. craccivora collected from Beni Suef was variable and differed slightly from the activity measured in the susceptible strain. Monitoring insecticide resistance among the three aphid populations was a proactive approach to detect any shift in insecticide efficiency. The possible occurrence of resistance in the cowpea aphid to the tested insecticides may be due to the higher activity of carboxylesterases. Further studies on the resistance mechanism to these insecticides are needed to provide insights in how to manage and delay the onset of the resistance and thus prolong the performance of insecticides against A. craccivora.     

一株大豆内生拮抗细菌XZ-2的分离和分子鉴定

从大豆植株中分离到一株具有广谱抗菌活性的拮抗细菌,命名为XZ-2。抑菌活性测定显示XZ-2对甘薯黑斑病菌、大蒜叶枯病菌等8种植物病原真菌均具有较强的抑菌活性,抑菌直径均在20 mm以上。采用分光光度法绘制了XZ-2的生长曲线,结果显示该菌株在LB液体摇培12 h时基本达到对数生长期,该生长曲线的绘制为后期XZ-2的活性物质发酵条件优化奠定了基础。通过PCR扩增16Sr DNA片段并采用MEGA 6.0构建系统发育进化树,初步将该菌株鉴定为多粘类芽孢杆菌。     

3,5-二硝基水杨酸法与酶法测定甜玉米还原糖和蔗糖含量的比较

针对目前测定甜玉米还原糖和蔗糖含量的3,5-二硝基水杨酸法和酶法进行比较研究,分析两者在育种中的应用。结果显示:酶法的测量准确度比3,5-二硝基水杨酸法高,且其稳定性和重现性都优于3,5-二硝基水杨酸法。通过两种方法对10个甜玉米杂交种种子的还原糖和蔗糖含量的测定,证实了两种方法在甜玉米还原糖和蔗糖含量的检测上是一致的。从育种实践考虑,酶法适于样品数较小的检测,3,5-二硝基水杨酸法更适合于大批量样品的检测。