柑桔酸性转化酶基因家族成员的克隆及特性分析

分析植物液泡和细胞壁酸性转化酶基因的保守区序列 ,设计 2对PCR引物 ,以温州蜜柑基因组DNA为模板 ,采用PCR方法扩增出长分别为 74 1bp(A)和 5 2 4bp(B)的 2个DNA片段 ,克隆入pUCm T载体测序 ,序列已在GenBank中登记 (登记号分别为AY0 2 94 81和AF332 881)。在GenBank中进行同源性检索 ,结果表明片段A编码的氨基酸与植物液泡酸性转化酶同源性较高 ,与胡萝卜、马铃薯和番茄同源性分别为 79%、79%和 78%。片段B编码的氨基酸序列与草莓、拟南芥和豌豆细胞壁转化酶同源性分别为 78%、78%和 77%。推测A和B分别定位于液泡和细胞壁。运用Clustalx和Bioedit软件包对已获得的柑桔转化酶基因家族的 7个基因序列进行分析 ,结果表明 7个基因分属转化酶基因家族的 4种类型。推测A和B为 2个新成员 ,分别属于柑桔液泡酸性转化酶基因Ⅱ型(CUAI2 )和细胞壁酸性转化酶基因Ⅱ型 (CUCWI)。Southern杂交结果表明 ,CUCWI和CSCWI 2个探针有时会出现较弱的杂交干扰信号 ,可采用CSCWI与CUCWI同源性较低的另一段序列制作探针进行杂交 ,也可通过缩短杂交时间、提高杂交和洗膜的温度以及降低洗膜SSC的浓度等措施来降低 ;其它成员之间无干扰信号产生     

铁铝氧化物和高岭石对转化酶的吸附及活性的影响

Experiments were conducted to study the influences of synthetic bayerite, non-crystalline aluminum oxide (N-AlOH), goethite, non-crystalline iron oxide (N-FeOH) and kaolinite on the adsorption, activity, kinetics and thermal stability of invertase. Adsorption of invertase on iron, aluminum oxides fitted Langmuir equation. The amount of invertase held on the minerals followed the sequence kaolinite > goethite > N-AlOH > bayerite > N-FeOH. No correlation was found between enzyme adsorption and the specific surface area of minerals examined. The differences in the surface structure of minerals and the arrangement of enzymatic molecules on mineral surfaces led to the different capacities of minerals for enzyme adsorption. The adsorption of invertase on bayerite, N-AlOH, goethite, N-FeOH and kaolinite was differently affected by pH. The order for the activity of invertase adsorbed on minerals was N-FeOH > N-AlOH > bayerite > reak goethite > kaolinite. The inhibition effect of minerals on enzyme activity was kaolinite > crystalline oxides > non-crystalline oxides. The pH optimum of iron oxide- and aluminum oxide-invertase complexes was similar to that of free enzyme (pH 4.0), whereas the pH optimum of kaolinite-invertase complex was one pH unit higher than that of free enzyme. The affinity to substrate and the maximum reaction velocity as well as the thermal stability of combined invertase were lower than those of the free enzyme.     

No‐tillage with half‐amount residue retention enhances microbial functional diversity,enzyme activity and glomalin‐related soil protein content within soil aggregates

A 3‐year field tillage and residue management experiment established in North China was used to analyse topsoil (0–15 cm) aggregation, and microbial functional diversity, enzyme activity and glomalin‐related soil protein (GRSP) content within aggregates. Compared with conventional tillage (CT), no‐tillage (NT) alone significantly (P < 0.05) increased organic C contents in 50–250 and <2 μm aggregates and decreased the proportion of C accumulated by 2–50 μm aggregates and microbial functional diversity indices in <2 μm aggregates. Regardless of tillage practice, both half‐amount (C50) and full (C100) residue retention tended to increase organic C and GRSP contents, or dehydrogenase and invertase activities, in certain aggregates. Under CT, a poorer performance of C50 than C100 was observed in maintaining Shannon index (H′) and Simpson index (D) in >250 and <2 μm aggregates, and also McIntosh index (U) in <2 μm aggregates, owing to insufficient residue and possible decreases in the distribution of decomposer micro‐organisms. Under NT, however, C50 was more effective than C100 in maintaining/elevating H′, D and U in all soil aggregates except for 50–250 μm, suggesting that surplus residue may induce worse soil conditions, decreasing heterotrophic microbial activities. Thus, NT with half‐amount residue retention improved soil physical–chemical–biological properties and could be a useful management practice in North China.     

沙棘根际土壤有机质特征及转化酶活性的初步研究

文章对沙棘根际和非根际的有机质特征和转化酶活性进行了初步的比较研究。结果表明，沙棘根际土壤有机质和活性有机质含量、可浸提腐殖酸及胡敏酸和富里酸含量、胡敏酸E4／E6比值都高于非根际土壤。根际土壤转化酶活性是非根际土壤的1．73倍。说明根际是树木和土壤相互作用最直接的区域，根际效应是树木改良土壤的重要机理。     

甘薯块根发育过程中关键蔗糖分解酶及其基因家族成员的鉴定

蔗糖分解酶在马铃薯和胡萝卜等块根/块茎的生长发育中发挥着重要作用,但在甘薯块根发育中的作用机制尚不清楚。本研究以2个在块根数量和鲜重上存在显著差异的甘薯品种‘高系14'及其突变体为材料,对其不同发育时期（30、60、90、120 d）块根中的可溶性糖（蔗糖、葡萄糖和果糖）、淀粉含量、蔗糖分解酶活性以及相关基因家族成员的表达水平进行测定,以明确调控块根数量和大小的关键蔗糖分解酶及主要基因家族成员。结果表明：（1）阐明了4种蔗糖分解酶活性在薯块根发育过程中的变化规律,细胞质转化酶（CIN）和液泡转化酶（VIN）活性的整体变化趋势呈现‘u'形曲线,即块根发育早期、晚期活性相对较高,发育中期最低;细胞壁转化酶（CWIN）和蔗糖合成酶（Sus）活性整体变化趋势呈‘n'形曲线,与前者正好相反,即在块根发育早期、晚期较低,发育中期最高。（2）和突变体相比,具有较高块根数量和鲜重的‘高系14'在块根发育早期（30 d）具有较高的Sus和CIN活性,而高Sus和CIN活性可以促进蔗糖由叶片向块根转运,从而提高块根中的淀粉、蔗糖和葡萄糖含量,最终为块根的生长发育提供能量和碳骨架以增加块根数量和鲜重。（3）从甘薯基因组中共鉴定出9个Sus基因家族成员和12个CIN基因家族成员,其中有1个Sus基因（IbSus6）和5个CIN基因（IbCIN4、IbCIN6、IbCIN8、IbCIN10和IbCIN11）的表达水平在30 d时表现为‘高系14'显著高于突变体,同时IbSus6和IbCIN4、IbCIN8、IbCIN10、IbCIN11分别为30 d块根中表达的主要Sus和CIN基因家族成员,因此上述1个Sus基因家族成员和4个CIN基因家族成员可能是调控甘薯块根发育的主要蔗糖分解酶基因。总之,Sus和CIN在甘薯块根早期发育中发挥着重要作用,其关键基因家族成员的阐明可为优异甘薯新品种的选育提供理论依据。     

施氮量对两个粒型小麦品种籽粒胚乳细胞增殖及糖含量的影响

为了解小麦胚乳细胞增殖、糖代谢在粒型间的差异及氮营养的调控效应,2009-2010年度以大粒型品种郑育麦9987和小粒型品种郑麦004为材料,研究了施氮量对小麦籽粒胚乳细胞增殖、细胞壁转化酶活性及糖含量的影响。结果表明,郑育麦9987胚乳细胞数(11.92×105)较郑麦004(10.73×105)多11.1%。施氮量影响籽粒胚乳细胞数,其中郑育麦9987低氮处理(150kg·hm-2)的胚乳细胞数目较高氮处理(300kg·hm-2)增加29.1%,而郑麦004则表现为高氮处理较低氮处理增加44.7%。两品种籽粒细胞壁转化酶活性均于花后5~8d达到峰值,花后20d则降到较低水平;籽粒可溶性糖和蔗糖含量均随灌浆进程整体呈先上升后下降趋势,在花后25d有一个小波峰。增施氮肥可增加籽粒细胞壁转化酶活性,提高后期籽粒可溶性糖和蔗糖含量。表明增施氮肥有利于小麦籽粒胚乳细胞增殖、光合产物的积累与运转,从而提高了粒重。     

Untersuchungen zur Assimilatakkumulation im Weizenblatt nach Schwarzrostinfektion

Accumulation of assimilates in stem-rust-infected wheat leaves
Using wheat plants (cv. 'Prelude'), the effects of stem-rust infection on photosynthesis and assimilate partitioning were studied with the aid of the short-lived carbon isotope ¹¹C. Photosynthesis of strongly infected leaf parts declined by about 30 % within 1 day. This rapid inhibition was caused by the formation of appressoria and the growth of infection tubes into the stomata, leading to a reduction in gas exchange. Further decline of photosynthesis to 5–10 % was accompanied by the degradation of chlorophyll. Nevertheless, during sporulation, the dry weight of the infected leaf area increased by about 50 %. With the use of ¹¹C, the reduced export of photosynthate from the infected leaf part and the capture of labelled compounds from the apical leaf section were quantified. The enhanced storage of assimilates in stem-rust-infected leaves can be attributed to the increase in invertase activity.     

兰州百合鳞茎可溶性酸性转化酶活性检测体系的建立

 为深入研究百合鳞茎蔗糖代谢生理机制,以兰州百合（Lilium davidii var. unicolor）鳞茎外层鳞片为试材,研究了不同种类和pH值的缓冲液、反应时间和反应温度对可溶性酸性转化酶（Soluble acid invertase,SAI,EC 3.2.1.26）活性的影响,建立了SAI活性的最佳检测体系。结果表明,最适提取缓冲液为pH 8.0的Hepes-NaOH;最佳反应缓冲液为pH 4.8的HAc-K3PO4,最适反应温度为40 ℃,最适反应时间为30 min。     

不同激素对红花子叶外植体过氧化物酶、酯酶同工酶和过氧化物酶、转化酶活性的影响

在 IAA、2,4-D 和 BA 作用下,红花子叶外植体过氧化物酶活性发生变化而同工酶谱带无显著改变,2,4-D 和 BA 显示对酶活性的累加促进效果。不同激素均可诱导红花子叶产生特异的酯酶同工酶酶谱。转化酶与激素所诱导的子叶外植体膨胀生长有密切联系。     

Dynamic Characteristics of Soil Sucrase and Amylase Activities during Vegetation Degradation in Gahai Wetland北大核心CSCD

In order to study the activities and dynamic characteristics of soil invertase and amylase during the process of vegetation degradation (Non-degradation, Light degradation,Moderate degradation, and Heavy degradation),the filed experiments were conducted in marsh meadow of Gahai wetland in Gannan state,Gansu province. The main results were as following: the average activity values of invertase and amylase activities in 0~100 cm layers varied in different stages of vegetation degradation of marsh meadow, following the order of CK (Non-degradation) > SD (Light degradation) > MD (Moderate degradation) > HD (Heavy degradation). With the increasing of soil layers,the activity of invertase significantly decreased (P<0.05),while the activity of amylase decreased fluctuantly. The activities of enzymes in the surface soil (0~20 cm) accounted for more than 60% of the total activities of each enzyme. The activities of invertase and amylase showed the obvious dynamic characteristics with time in all stages of vegetation degradation. The average activity of invertase in different months was the highest in June (47.07 mg·g-1) and the lowest in August (22.69 mg·g-1). The average activity of amylase was the highest in July,85.18 mg·g-1,while the activity values in other months were low. Correlation analysis showed that soil organic carbon was significantly correlated with invertase and amylase (P<0.01),and soil water content was only significantly correlated with invertase (P<0.01). The results showed that the vegetation degradation in Gahai wetland significantly affects soil invertase and amylase activities,reduces the efficiency of enzymatic reaction,and leads to the deterioration of wetland soil environment. © 2019 Ｊｏｕｒｎａｌ ｏｆ Ｒｅｆｒｉｇｅｒａｔｉｏｎ. All rights reserved.