一个潜在催化莱鲍迪D苷合成的新型糖基转移酶

尿苷二磷酸糖基转移酶(uridine diphosphate glycosyltransferases,UGTs)催化糖基转移反应,与植物次生代谢密切相关。本研究根据甜叶菊(Stevia rebaudiana)转录组数据库,克隆到一个催化莱鲍迪D苷(rebaudioside D,RD)合成的新型糖基转移酶候选基因,对其开展生物信息学分析。结果表明,该基因开放阅读框长1380 bp,编码459个氨基酸,等电点(pI)预测为5.54,理论分子量约49.66 kD,系统发育分析表明该基因与向日葵中的UGT89A2同源,故将其命名为SrUGT89A2。构建pET28a-SrUGT89A2原核表达载体,并在大肠杆菌(BL21(DE3))中诱导表达得到重组蛋白,HPLC检测表明粗酶液能催化甜叶菊提取液形成一个新的色谱峰,该峰保留时间与莱鲍迪D苷一致。经进一步纯化UGT89A2蛋白,添加不同甜菊糖苷标准品为催化底物,但未鉴定出该蛋白催化的具体糖苷。该潜在催化甜菊糖RD苷合成的新型糖基转移酶基因SrUGT89A2的发现,为RD苷的生物合成和甜菊糖苷的生物途径研究提供新的理论依据。     

蔬菜幼苗叶面积快速测定方法筛选与优化

为得到一种快速且准确测定蔬菜幼苗叶面积的方法,以番茄幼苗为试材,以方格法为对照,研究扫描分析法、叶面积仪法、打孔称重法测定叶面积的准确性、精确性与测定速度,然后采用黄瓜、辣椒叶片及纸片的面积对测定结果进行验证,最后基于不同扫描分辨率对筛选出的方法进行优化。结果表明,相比叶面积仪法和打孔称重法,扫描分析法测定结果更接近真实值,且精度更高,用时更短。扫描分析法不同分辨率下测定结果无显著差异,且扫描分辨率越低所需测定时间越短。扫描分辨率50 dpi下的扫描分析法可作为一种快速准确测定蔬菜幼苗叶面积的方法。     

辣椒种质资源形态学性状相关性、主成分与聚类分析

以全国各地搜集的205份辣椒种质为试材,观测28个形态性状,并对其进行相关性、主成分和聚类分析研究。结果表明:辣椒形态学性状间存在较高相关性,单株果数、叶形、株型、果形、茎绒毛、花梗着状、熟性、花柱色是最关键的性状。6个主成分的方差累积贡献率为50.653%,暗示这28个性状代表了更多的性状改良方向。主成分归纳为形态因子、产量因子、熟期因子、色彩因子,呼应聚类分析的主要区分依据果形、单株果数、熟期和果色,将205份种质分成7类,有效区分了不同的品种。说明在改良形态性状时,按育种目标的要求选取主要性状,同时也要注意与其它性状间的关联。     

Effect of Kechuanning on airway remodeling and protein level of p-ERK1/2 in lung tissues of asthmatic rats induced by virus

AIM:To investigate the effect of Kechuanning on airway remodeling and the protein level of p-ERK1/2 in lung tissues of asthmatic rats induced by virus. METHODS:The asthmatic rat model induced by respiratory syncytial virus was established. The experimental rats were divided into normal group, asthma model group, low dose (0.33 mL/kg), middle dose (3.0 mL/kg) and high dose (10 mL/kg) of Kechuanning groups, and PD98059 (3 mg/kg) group. The airway responsiveness of the rats was measured by animal ventilator. The pathological changes of the lung tissues were observed by HE staining. PAS staining and Masson staining were used to observe goblet epithelial cells metaplasia and airway collagen deposition. The expression of matrix metalloproteinases-9 (MMP-9) and tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) in the lung tissues of the rats was detected by immunohistochemical staining. The protein levels of ERK1/2 and p-ERK1/2 were determined by Western blot. RESULTS:Compared with model group, the airway responsiveness of the rats in middle dose and high dose of Kechuanning groups was significantly decreased (P<0.01), the injury of lung tissues was significantly decreased, the goblet epithelial cells metaplasia and airway collagen deposition were significantly reduced (P<0.01), and the expression of MMP-9 and TIMP-1 in the lung tissues was also significantly decreased (P<0.01). In addition, the protein level of p-ERK1/2 in high dose of Kechuanning group was significantly decreased compared with model group (P<0.01). CONCLUSION:Kechuanning may treat asthma by regulating the expression of p-ERK1/2 in the lung tissues and improving the airway remodeling symptoms of asthmatic rats induced by virus.     

桑植县茶产业现状及发展对策

通过对桑植县茶产业现状进行实地考察调研,剖析桑植县茶产业发展的优势与不足,提出桑植县茶产业发展的对策措施。     

完善内地主产棉区棉花补贴机制的思考与建议——以山东省为例

总结了棉花目标价格补贴政策实施以来山东植棉区的实施进展、成效与问题。从国家棉花产业安全和山东省经济社会发展角度，分析了山东植棉区棉花生产的重要地位和保持补贴政策稳定性的重要意义。以山东为例，提出了内地主产棉区棉花补贴政策改革建议。     

马铃薯晚疫病菌效应子PITG_16427.2的基因序列分析及功能验证

 马铃薯晚疫病的病原是致病疫霉菌(Phytophthora infestans),该病原菌在侵染马铃薯过程中分泌大量RxLR型效应子,但目前绝大多数RxLR效应子的功能和作用机制尚不明确。本研究成功克隆了致病疫霉菌的一个RxLR效应子PITG_16427.2,在本氏烟中瞬时表达PITG_16427.2,发现该效应子能够抑制6种激发子(INF1、PsojNIP、BAX、SIF2、Avh238、Avh241)激发的植物免疫反应。进一步研究发现,效应子PITG_16427.2在晚疫病菌侵染马铃薯早期上调表达。在15个致病疫霉菌株和3个同属菌株中克隆该基因,克隆到氨基酸序列一致性超过93%的同源基因,这些同源基因均能在本氏烟中抑制INF1和Avh241引起的HR,揭示了该效应子在病原卵菌中序列和功能的高度保守性。在本氏烟和马铃薯感病品种Désirée中瞬时表达PITG_16427.2,发现该效应子能够显著促进晚疫病菌的侵染。通过qRT-PCR方法检测发现,乙烯信号的相关基因ERF1显著上调,而水杨酸信号相关基因PR1b显著下调,表明PITG_16427.2在晚疫病菌侵染过程中可抑制寄主的SA信号途径,促进晚疫病菌侵染。因此,RxLR型效应子PITG_16427.2是致病疫霉菌中一个重要的侵染致病因子。     

海贝柱自动晾晒设备设计

针对当前传统人工晾晒海贝柱效率低下、容易造成污染等问题。设计了海贝柱自动晾晒机械,采用阶梯式晾晒生产线对湿海贝柱分级晾晒,多条晾晒生产线分时、高效的工作,机器上方设有遮板以保证晾晒作业不被污染而且不受天气条件的影响。整机采用stm32单片机、传感器等技术实现自动化作业,机器控制系统监测到生产线发生故障或海贝柱出现堆积时会立即停止工作以便于检查故障、减少损失。试验结果表明:机械晾晒不会造成海贝柱内的营养成分的流失,机械晾晒海贝柱的干基含水率达到30%仅需12h,机械晾晒海贝柱能节省大量的干燥时间,显著的提高了晾晒效率,避免了人工晾晒过程导致的二次污染和损失。该研究为海贝柱自动晾晒机械的研制提供了技术参考。     

减氮控水对日光温室番茄-小型西瓜产量、品质及氮素利用的影响

研究日光温室滴灌施肥条件下,不同水肥组合对作物产量、品质和氮素利用的影响,综合评价优化水肥和常规水肥管理的调控效应。田间试验设置4个水氮处理,包括不施氮+常规灌溉(N_0+FI)、常规施氮+常规灌溉(FT+FI)、优化施氮+常规灌溉(OPT+FI)、优化施氮+优化灌溉(OPT+OI)。测定不同处理下秋冬茬番茄-春茬小型西瓜的产量、品质以及根、茎、叶、果实氮素吸收量。结果表明:优化水氮处理(OPT+OI)番茄产量和果实可溶性糖、可滴定酸和Vc含量与农户常规水氮处理(FT+FI)无显著差异,但番茄果实的硝酸盐含量降低了66.3%(P0.05);灌溉量相同时,减氮40%处理(OPT+FI)的小型西瓜产量相比常规施氮处理(FT+FI)提高了13.1%;OPT+OI处理果实的可溶性糖、可滴定酸和Vc含量较对照处理(N_0+FI)均显著提高。不同水肥处理下,两季作物氮在各器官的累积量均表现为果实叶茎根。随着番茄的生长,果实和茎的氮素携出量占总携出量的比例分别由62.4%和5.9%增加至67.1%和6.3%,而根和叶中氮素携出量降低,OPT+OI处理果实氮素携出所占比例增量最大,促进了营养器官中的氮素向果实中转运。在当前日光温室栽培条件下,适当优化施氮量和灌溉量既可以保证作物产量和氮素吸收,同时提高果实的品质。     

Detection of seven phytohormones in peanut tissues by ultra-highperformance liquid chromatography-triple quadrupole tandem mass spectrometry

Development of highly sensitive and reliable method for detection of phytohormones is of great significance to study plant hormones and agricultural production.In this study,an ultra-high-performance liquid chromatography-mass spectrometry/mass spectrometry method was established for separation and quantification of trans-zeatin,trans-zeatin riboside,gibberellin A_3,indol-3-acetic acid,salicylic acid,abscisic acid,and jasmonic acid(JA) without any label.The sepa ration was performed on an Agilent Explus Plus C18 column by using methanol and water as mobile phases with gradient elution.The target compounds were confirmed and quantified by mass spectrum via positive electrospray ionization for trans-zeatin,transzeatin riboside,indole-3-acetic acid,and via negative electrospray ionization for gibberellin3,salicylic acid,abscisic acid,and JA.The limits of detection ranged from 0.0127 ng L~(-1) for gibberellin A_3(GA_3) to 33.26 ng L~(-1) for JA and were lower than the currently reported values in literature.The proposed method was applied for qualitative and quantitative analyses of phytohormones in peanut gynophores and pods.The recoveries of the spiked phytohormones ranged from 80.20 to102.56%.The contents of seven endogenous hormones varied specifically in different development stages of peanuts.This study provides a highly sensitive and selective detection method for hormones and elucidates the growth and development of the gynophore and peanut fruit,which are controlled by seven endogenous hormones.