基于多反应监测质谱技术的水稻叶片蛋白绝对定量方法

【目的】建立水稻叶片蛋白的多反应监测(MRM)质谱绝对定量方法。【方法】水稻叶片蛋白经含1.0%十二烷基硫酸钠(SDS)的磷酸盐(PBS)缓冲液提取,丙酮沉淀除杂纯化、胰蛋白酶消化,酶解液经液相色谱分离,MRM质谱监测,外标法定量。【结果】向提取缓冲液中加入1.0%SDS可增强水稻叶片中16种靶蛋白和总蛋白质的提取效果;不同有机试剂处理,总蛋白质沉淀量存在显著差异(P<0.05),沉淀能力从强到弱依次为乙腈>丙酮>异丙醇>甲醇>乙醇;对于16种目标蛋白,总体以丙酮沉淀效果最好,其次是异丙醇和乙腈,甲醇和乙醇效果较差。该方法线性范围均达到3个数量级,定量限为0.1~2.5 nmol/L,灌浆期16种水稻叶片蛋白质含量为6.0~2818.1μg/g,相对标准偏差均小于14%。【结论】SDS可显著提高水稻叶片蛋白提取效果,采用丙酮或乙腈可获得较好的蛋白沉淀效果,但不同蛋白质略有差异。结合MRM质谱监测技术可实现水稻叶片蛋白的绝对定量,方法线性范围宽、敏度高、重复性好。     

杂交稻低播量精量播种育秧及机插取秧特性

【目的】明确杂交稻70 g以下低播量精量穴播和条播对育秧效果及机插特性的影响,突破生产中杂交稻机插的技术瓶颈。【方法】以中浙优1号和甬优1540两个杂交稻品种为材料,利用机插标准9寸盘,设置机械穴播和条播两个精量播种方式,并以机械流水线撒播为对照,穴播规格为16(纵向)×34(横向)穴,条播为纵向16条,以穴播5粒、3粒及2粒的播种量进行播种试验。考查了低播量下不同播种方式对秧苗生长影响及配套取秧效果【结果】1)低播量下不同播种方式对秧苗成苗率的影响不大。2)降低播种量提高秧苗生长一致性,且穴播和条播秧苗生长一致性好于撒播。3)与撒播相比,精量穴播和条播能够在低播量下提高秧苗根系盘结力和成毯性,中浙优1号和甬优1540根系盘结力比撒播平均高75.4%和81.0%,播量每穴3粒时即能有效成毯,穴播和条播差异不大。4)精量穴播和条播能够显著降低低播量下机插漏秧率,中浙优1号和甬优1540机插漏秧率平均分别比撒播低76.3%和74.6%,穴播和条播下,两个品种每穴播量3粒的漏秧率均在1%以下,与撒播相比降幅在10个百分点以上。5)精量穴播和条播机插取秧苗数均匀度比撒播要好,两个品种预期取秧2~5苗(5粒)、1~3苗(3粒)和1~2苗(2粒)比例均达80%以上,中浙优1号和甬优1540机插苗数均匀度平均比撒播高121.2%和67.0%,其中,穴播机插取秧苗数均匀度及预期取秧苗数比例高于条播。【结论】精量穴播和条播可以解决目前杂交稻机插用种量大、漏秧率高和机插取秧苗数均匀度差的问题。     

四端法土壤电导率传感器恒流源设计与试验

土壤信息对于指导农业生产有着极为重要的作用，土壤电导率反映了土壤含水量、盐分、粘粒含量和类型等土壤信息，准确获取土壤电导率对于实现农业精细化生产意义重大。在各种土壤电导率测量方法中，四端法因其成本低、精度高、测量快速和操作简便而大量应用于实际测量；恒流源是四端法测量仪器的重要组成部分，其性能直接决定着测量仪器的精度以及测量范围。本文对比了3种恒流源对测量仪器测量性能的影响，发现采用Howland恒流源的四端法测量仪器高电导率测量能力强而低电导率测量范围较小，测量精度最高；采用改进型Howland恒流源的四端法测量仪器低电导率测量范围有所扩大，测量精度良好；采用基于差动放大器的恒流源的电导率仪低电导率测量范围最大，高电导率测量范围较优，测量精度良好。     

提取方法对库尔勒香梨果实表皮蜡质提取效果的影响

以库尔勒香梨为试材，三氯甲烷和正己烷为提取剂，采用常温法、加热法和常温加热组合法提取香梨果实表皮蜡质，探讨香梨果皮蜡质的提取方法，为研究香梨果皮蜡质结构及成分提供基础。结果表明：以三氯甲烷为提取剂的效果明显优于正己烷，采取常温结合加热法提取表皮蜡质的含量最高，常温法提取的蜡质含量最低。扫描电镜观察结果表明：采用常温加热结合法，以三氯甲烷作溶剂可充分提取香梨表皮蜡质，而正己烷提取果实表皮呈不明显网状结构，仍有晶体残留；采用加热法，以三氯甲烷提取香梨果实表皮蜡质后，表皮蜡质有小部分残留，而正己烷提取后残留晶体较三氯甲烷提取后多，有不规则的小粒状晶体残存；用常温法则两种溶剂均不能将香梨表皮蜡质提取干净。综合提取蜡质含量和扫描电镜分析的结果得出，以三氯甲烷为溶剂、采用常温加热结合法是有效且易行的提取库尔勒香梨表皮蜡质的方法。     

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。     

Age estimation of different tree species using a special kind of an electrically recording resistance drill

The determination of tree age is an important issue for urban green planning, forestry and dendrology; finding non-destructive and quasi-non-destructive methods for this purpose is of great theoretical and practical importance. The resistance drilling method is quasi-non-destructive because the average diameter of an opening that remains after drilling does not exceed 3 mm. Do electrically recording resistance drills allow precise assessment of tree age? The aim of this study is to assess the accuracy of determining the number of tree rings based on an examination of this special kind of drilling resistance profiles for three tree species, the pine Pinus sylvestris L., the oak Quercus robur L., and the birch Betula pendula ROTH. In 2015 and 2016, 15 pine trees, 15 oak trees, and 15 birch trees were randomly selected. For each studied tree, a measurement was conducted using the electrically recording resistance drill IML-Resi E400 with a flat-tipped 1.5/3 mm steel needle (research sample), and an increment core was taken (reference sample). The drill used was not a real Resistograph^®. The analysis of the E400-profiles underestimated the number of tree rings; the mean bias error (MBE) values were –6.5, –2.5, and –6.0 years for pine, oak, and birch, respectively. The proportion of investigated trees with less than five years difference between the research and reference samples varied from 38.4 (birch) to 66.7 (oak) percent. The accuracy of tree age determination was lowest for birch and highest for oak. The binomial generalised linear model (GLM) revealed that the most accurate tree age assessments were obtained from tree rings wider than 2 mm. The measurements clearly showed that the electrically recording resistance drill IML-Resi E400 enables a quick, although approximate, tree age assessment. Future research should concentrate on electronically regulating and recording drills, providing a higher spatial and signal resolution, and a stronger correlation to wood density.     

Beaver dams,streamflow complexity,and the distribution of a rare minnow,Lepidomeda copei

Freshwater fishes are threatened globally, and often too little is known about threatened species to effectively guide their conservation. Habitat complexity is linked to fish species diversity and persistence, and degraded streams often lack habitat complexity. Beaver Castor spp., in turn, have been used to restore streams and increase habitat complexity. The northern leatherside chub Lepidomeda copei is a rare, small‐bodied, drift‐feeding minnow that has anecdotally been observed to use complex habitats associated with beaver dams in the western United States. To investigate this anecdote, we conducted fish and habitat surveys, the latter focusing on quantifying habitat complexity, in a sub‐basin of the Upper Snake River Basin in the USA. Complementary generalised linear model and path analyses revealed that northern leatherside chub occurred more often at sites with complex streamflows, and streamflows were more complex when beaver dams were present and pools were deeper. Northern leatherside chubs were also more likely to occur when temperatures were warmer, aquatic macrophytes were abundant and stream channels were narrow and deep. The linkage between chubs, complex streamflows and beaver dams needs to be evaluated more broadly to completely understand its role in the rangewide status of the species. However, it does suggests that increased use of beaver reintroductions and dam analogues for stream restoration could be a boon for the northern leatherside chub, but such efforts should be monitored to determine their effectiveness to help adapt beaver‐based restoration approaches to best benefit the species.     

Responses of Australian sea lions,Neophoca cinerea,to anthropogenic activities in the Perth metropolitan area,Western Australia

1. Tourist‐based activities, partly due to their rapid increase, have raised concerns regarding the impacts of anthropogenic activity on marine fauna. Documented effects on pinnipeds in proximity to humans include changes in behaviour, site use and potentially higher aggression levels towards people. Effects vary considerably between populations and sites, thus requiring separate assessment of human impacts on activity and energy budgets.
2. Responses of the endangered Australian sea lion, Neophoca cinerea, to human visitation were recorded from November 2013 through April 2014. Exposure levels and response types to anthropogenic activities were assessed at two easily accessible locations with different management schemes, Seal (landing prohibited) and Carnac (landing permitted) islands, Western Australia. Exposure levels were measured as both stimulus type (i.e. ‘People’, ‘Paddlers’, ‘Small’, ‘Medium’, and ‘Large vessels’, ‘Tour vessels’, and ‘Jet skies’), and people (‘Direct’, ‘Attract’, ‘Interact’, ‘View’, ‘Incidental’, ‘Water’, ‘Low‐level’), and vessel activities (‘Interact’, ‘Approach/Follow’, ‘Anchor noise’, ‘Engine noise’, ‘Close to beach’, ‘Moderate/Fast travel’, ‘Slow travel’, ‘Transit’, ‘Drift/At anchor’, ‘Aircraft noise’).
3. Exposure levels varied significantly between the islands in numbers, stimuli type, duration and minimum approach distances. The instantaneous behaviours of ‘Lift head’, ‘Interact’ and ‘Sit’ were the most frequent responses. ‘Aggressive’ and ‘Retreat’ responses, the highest disturbance levels measured, occurred on Carnac approximately once per day, but rarely on Seal Island. ‘Aggressive’ behaviour towards ‘People’ was observed only on Carnac Island and elicited only by ‘People’. ‘People’, ‘Tour vessels’, and scenic ‘Aircrafts’ on both islands as well as ‘Jet skis’ on Carnac Island had the highest probability of triggering responses. Owing to their relatively high visitation at Seal Island, ‘Paddle powered vessels’, followed by ‘Tour vessels’ elicited the highest number of responses, compared with ‘People’, ‘Small’, and ‘Medium vessels’ at Carnac Island. The majority of responses occurred when any stimulus type was at short‐range (≤10 m), and ‘People’ ‘Viewing’ N. cinerea elicited most. Vessels triggered more responses at larger ranges than ‘People’.
4. To limit close‐range access to N. cinerea, one possibility is to close the beach at Carnac Island to human visitation and increase the minimum approach distance by vessels and ‘People’ by installing marker buoys at least 15 m from the shore.

© 2016 The Authors. Aquatic Conservation: Marine and Freshwater Ecosystems published by John Wiley & Sons, Ltd.     

Habitats for the Atlantic sturgeons in Russia

1. The critically endangered Atlantic sturgeon, Acipenser sturio, inhabited the Russian section of the Baltic Sea basin until recent times. Eight rivers were indicated as their habitats in local literature.
2. The Atlantic surgeon disappeared in Russia because of overfishing, not because of habitat loss.
3. Nowadays, potential spawning grounds for sturgeons occur in six rivers of the Russian section of the Baltic Sea basin. The largest river (Neva) is not blocked by dams and the whole river provides the ecological requirements of sturgeons on a stretch 75 km long.
4. Restocking of other sturgeon species in Russia demonstrated that the release of reared fishes into the sea is ineffective, whereas the release into rivers could result in successful acclimatization.
5. Recent climatic changes are rather favourable for sturgeon restocking in the Baltic Sea area. The main problem for the restoration of sturgeon populations is the intensive fishery.