糯小麦杂交后代的基因型鉴定及Wx基因的效应研究

糯小麦在食品加工业、淀粉加工业及其它工业上有着重要用途，是近年来许多国家小麦研究的重要课题。国外糯小麦选育尚未突破高产与糯性相结合的难点，国内目前还没有培育出高蛋白强筋型的糯小麦品种，这在一定程度上与缺乏合适的育种方法和高效、实用的糯小麦分子标记辅助育种技术有关。国内外对Wx基因效应的研究主要利用缺体－四体系、重组自交系或近等基因系，还未见有利用遗传背景相同的BC5F2代回交改良群体的报道。 糯性位点近等基因系是小麦淀粉品质育种的重要材料，而我国目前还没有一套中国栽培小麦遗传背景的糯性位点近等基因系。为了选育部分糯小麦、全糯小麦和中国栽培小麦遗传背景的糯性位点近等基因系，我们利用Wx蛋白电泳和高效实用的分子标记技术体系来鉴定糯小麦杂交后代的基因型，结果证明该体系能有效地用于糯小麦的分子标记辅助育种。以中国春糯性位点全套近等基因系为研究材料，对小麦Wx基因的6个STS标记和1个CAPS标记进行了筛选，改良PCR扩增条件以及产物检测方式后，从这些标记中筛选出3个标记，包括鉴定Wx-A1、Wx-D1位点的2个共显性STS标记和Wx-B1位点的1个显性STS标记。利用上述3个分子标记从BC5F2 代回交改良群体中筛选出了8种Wx基因型，经卡方检验，其分离比符合3对基因的分离比例，其中基因型为aabbdd的植株有2株，直链淀粉含量分别为1.81%和0.82%，为全糯小麦；基因型为AAbbdd，aabbDD的部分糯性植株各有1株，直链淀粉含量分别为15.24%和17.57%。以上4株植株的农艺性状和品质性状接近回交亲本“川育12”，并明显优于全糯材料“98Y1441”，表明采用回交法与Wx基因分子标记辅助选择相结合，有助于培育高产、优质的全糯和部分糯小麦。同时，本研究中建立的分子标记技术体系，也为选育具有中国栽培小麦遗传背景的糯性位点近等基因系奠定了基础。 在基因型鉴定的基础上，利用糯小麦杂交后代BC5F2代回交改良群体研究了各基因缺失降低直链淀粉含量的效果和各基因合成直链淀粉的能力，以及直链淀粉含量与农艺性状、品质性状、淀粉糊化特性等的相关性。缺失不同Wx基因的8种基因型，直链淀粉含量差异显著。研究单缺失基因型发现，减少效应最大的是Wx-B1b基因，Wx-B1b和Wx-D1b基因没有显著差异，减少效应最小的是Wx-A1b基因。研究双缺失基因型发现，Wx基因合成直链淀粉的能力，Wx-B1a基因最高，Wx-A1a基因最低，而Wx-B1a和Wx-D1a基因差异很小；直链淀粉含量与株高、穗长、小穗数、穗粒数、千粒重等农艺性状相关不显著，表明淀粉品质育种可以与高产育种实现有机结合；直链淀粉含量与SDS-沉降值呈显著负相关(r=-0.726)，说明直链淀粉含量降低在一定程度上有利于提高小麦营养与加工品质，这一结果至今未见有文献报道；全糯类型的淀粉糊化特性与其他类型显著不同，具有最高的峰值粘度和稀懈值，最低的低谷粘度、最终粘度、反弹值、峰值时间、糊化温度、起始糊化温度，表明糯小麦淀粉在食品和工业上具有特殊用途；稀懈值与直链淀粉含量呈极显著负相关(r=-0.969)，其他粘度参数与直链淀粉含量呈显著正相关(最终粘度r=0.797，低谷粘度r=0.910、反弹值r=0.954、峰值时间r=0.970、糊化温度r=0.962、起始糊化温度r=0.932)。以上结论可为糯小麦品种选育和淀粉品质改良提供理论依据。 Waxy wheat is very important in food processing industry, starch processing industry and the other industries, so it is a focus of wheat research in many countries these years. Foreign wheat breeders have not conquered the difficulty of high yield combined with waxy character, and there is no waxy wheat variety with high protein and strong gluten in China at present，all of which were caused by lacking proper breeding methods and effective, applied molecular markers-assisted selection technique at a certain extent. Until now, research about the effect of waxy genes mainly depended on nullisomic-tetrasomic lines, recombinant lines or near-isogenic lines, and it is lacking the reports of using improved BC5F2 backcross progenies under the common genetic background. Near-isogenic lines at the Wx loci are important materials for wheat starch quality breeding. However, there are no such lines under the Chinese cultivated wheat genetic background. To develop partial waxy wheats, waxy wheats and near-isogenic lines at the Wx loci under the Chinese cultivated wheat genetic background, we use SDS-PAGE of waxy proteins and effective, applied molecular marker-assisted selection technical system to identify the genotype of waxy wheat’s progenies. The results indicated that such a system is applicable in waxy wheat’s molecular marker-assisted selection effectively. A series of Chinese Spring Wx loci near-isogenic lines were used to identify the specific bands of six STS markers and one CAPS marker of Wx genes. After optimizing PCR amplification and separating of PCR products, three co-dominant and dominant STS-markers were identified at the Wx-A1, Wx-D1 and Wx-B1 loci, respectively, which were used to identify the genotype of waxy wheat’s progenies. Eight Wx genotypes were developed from the improved BC5F2 backcross progenies, which follows Mendelian segregation. Among them, there were two aabbdd waxy plants whose amylose content in the BC5F3 seeds were 1.81% and 0.82%, respectively. In addition, there were partial waxy plants (AAbbdd and aabbDD) whose amylose content in the BC5F3 seeds were 15.24 % and 17.57%, respectively. Most agronomic and quality traits of these four plants resembled those of the recurrent parent “Chuanyu 12”, and superior to waxy wheat parent “98Y1441”. This shows that backcross approach in combination with molecular marker-assisted selection of waxy genes is helpful to develop partial and full waxy wheat with good traits in the waxy wheat breeding program. At the same time, molecular marker-assisted selection technical system in this paper, also establish the base for breeding the near-isogenic lines at the Wx loci under the Chinese cultivated wheat genetic background. According to the results of genotype identification, we use waxy wheat’s improved BC5F2 backcross progenies to verify the effects of null alleles on reducing amylose content and determine the amylose synthesis capacity of each Wx gene independently, and investigate the relativity among amylose content with agonomic traits, quality traits, starch pasting properties respectively. There was significant difference in amylose content of the eight genotypes carrying different null Wx alleles. The reducing effect of the single null alleles was the most significant in Wx-B1b, followed by Wx-D1b and Wx-A1b, and there was no significant difference between Wx-B1b and Wx-D1b. The results of the double null lines further demonstrated that for the capacity of amylose synthesis, Wx-B1a was the highest, followed by Wx-D1a and Wx-A1a, and there was no significant difference between Wx-B1a and Wx-D1a. Amylose contents of the eight genotypes were not significantly correlated with plant height, spike length, spikelets per spike, grains per spike, 1000-grain weight, which showed that starch quality breeding could integrate with high yield breeding. Amylose contents of the eight genotypes were negatively significantly correlated with SDS-sedimentation value(r=-0.726), which suggested that reduction in amylose content is propitious to improve quality at a certain extent. Starch pasting properties of the full waxy type was significantly different from the other seven types, with the highest peak viscosity and breakdown, and lowest valley viscosity, final viscosity, setback, peak time, pasting temperature and starting pasting temperature. It indicated that waxy wheat starch has special use in food and industry. Breakdown was negatively significantly correlated with amylose contents (r=-0.969), and the other parameters were positively significantly correlated with amylose contents (r=0.797 for final viscosity, r=0.910 for trough viscosity, r =0.954 for setback, r =0.970 for peak time, r=0.962 for pasting temperature and r=0.932 for starting pasting temperature ). The results of the study are very useful for waxy wheat variety breeding and starch quality improvement.

Detection of K-ras exon 1 mutations by constant denaturant capillary electrophoresis

Among various mutation detection methods, constant denaturant capillary electrophoresis (CDCE) is one of the most common techniques for rapid identification of known or unknown mutations. In this report, a CDCE analysis method with homemade linear polyacrylamide (LPA) kit was developed on ABI 310 genetic analyzer, the effect and relationship of various denaturing factors in CDCE analysis were investigated and K-ras gene mutations of 31 coloerctal cancer patients were detected. Results indicate that, with the increase of chemical danaturant concentration, the optimum temperature was lowered, and when the concentration of urea (formamide) was higher than 7 M (40%), the homoduplex and heteroduplex of mutant samples were separated with difficulty. Detection results of K-ras gene in colorectal samples indicated that mutations were present in eight (26%) of 31 patients; most mutations were localized in codon 12, which is thought to be a critical step and plays an important role in human colorectal carcinogenesisas. Copyright (C) 2004 John Wiley Sons, Ltd.

浅论实现基于MapX的灌溉信息系统的关键技术

现代化的灌溉用水管理是实现农业高效用水的重要措施之一,它可实现灌溉水资源的合理配置和灌溉系统的优化调度,达到节水增产目的,使有限的水资源获得最大的效益。采用手工方式进行灌溉用水管理,难以及时提供水资源利用最优化方案,因此急需研制出具有网络通信功能的可视化操作界面的灌溉信息管理系统,为灌区水资源评价和利用、规划等工作奠定良好的数据基础。GIS不仅具有强大的空间数据管理功能,还具有丰富的多元地学数据分析处理能力和直观的图形显示效果,因此利用GIS进行灌区的管理与建设将会有效提高灌区的工作效率和经济效益。笔者在实践调研的基础上,根据灌溉信息管理系统的功能规划及结构设计,利用MapX组件工具开发了陕西省冯家山灌区灌溉信息系统,对提高灌溉用水的管理效率进行了实例尝试,并着重对下列关键技术的运用进行了实践与思考:(1)阐述了利用可视化编程工具结合数据库管理技术、地理信息系统组件技术、网络通信技术等进行系统开发的过程;(2)着重介绍了基于MapX组件的空间数据库的建立与关联技术、灌溉管理信息的网络通信实现技术;(3)最后对在灌溉管理工作中运用GIS技术进行科学管理等相关问题进行了有益的探讨。

基于CorelDRAW软件的小流域模型雨滴测量试验研究

降雨雨谱的观测,是了解降雨特性的一项重要工作。修建小流域模型,是对小流域进行各种试验研究的有效手段。该文基于相似率要求,在模型降雨雨滴较小,雨谱参数难以测定问题的基础上,运用CorelDRAW软件与传统色斑法相结合,探索出一种新的测量与数据处理方法,并得出不同情况下模型的雨谱特性。该法提高了测量精度,并减少了工作量。

Interfacial characteristics of the self-assembly system of poly-L-lysine 3-mercaptopropionic acid gold electrode

The interfacial characteristics of poly-L-lysine (PL) attached on self-assembled monolayers (SAMs) of 3-mercaptopropionic acid (MPA) were studied by an electrochemical method. The results indicated that PL\MPA layer inhibited partly the diffusion process of redox species in solution, and the electrode surface behaved like a microelectrode array. Its permeation effect was also strongly affected by Mg2+. The more Mg2+ ions were added into the electrolyte solution, the greater the difficulty with which the electron transfer of potassium ferricyanide took place. The three different conformations of PL on the electrode surface had different influences on the electron transfer processes of ferricyanide. PL in random coil state hindered most strongly the electron transfer behavior of ferricyanide,while the alpha-helical PL had nearly no effect and the effect of the beta-sheet state PL was intermediate of these. (C) 1997 Elsevier Science S.A.

Cultivation of the intertidal brown alga Hizikia fusiformis (Harvey) Okamura: mass production of zygote-derived seedlings under commercial cultivation conditions, a case study experience

Mariculture of the brown alga Hizikia fusiformis (Harvey) Okamura as an export-oriented human food has been there more for than 20 years in China. It is now one of the five major farmed algal species along the Chinese coast. Stable and sufficient supply of young seedlings for scaling up the cultivation has been a problem throughout the farming history of this species due to the unique dioecious life cycle and relatively short time window of sexual reproduction in nature. These two factors led to a practical difficulty in obtaining zygotes at identical developmental stage in viable amounts for seedling production. A key solution to this problem is to control the synchronization of the receptacle development and to realize the simultaneous discharge of male and female gametes, such that the fertilization rate could be greatly enhanced. Focusing on one of the farmed populations in this report, we present our results on mass production of seedlings using the synchronization technique on a large scale performed in 2007. Totally 5.5 hundred million embryos were obtained from 100 kg female sporophytes. The seedlings were raised up to 3.5 mm in length in greenhouse tanks over a month and were further grown in open sea for over 3 months at two experimental sites. The success of mass production of seedlings in this alga helped to lay the basis for future trials in other species in the genus of Sargassum that have identical life cycle.

Improvement of polyploidy induction in Eriocheir sinensis

An incubating temperature of 15 degreesC is used to induce triploidy in Etiocheir sinensis through inhibition of the release of polar body H, and that of 18 degreesC to induce tetraploidy through inhibition of the first cleavage. Flow cytometry is used to identify the ploidy in different developmental stages. For induction of triploidy in fertilized eggs in vitro, the highest induction rate observed in blastula by cytochalasin B, 6-DMAP and KCI is 49.1%, 51.7% and 77.5%, respectively. In the KCI treatment of pregnant crabs with the fertilized eggs, the highest triploid induction rate observed in the zoea is 85.3%. For induction of tetraploidy, the highest induction rate observed in the blastula by cytochaslasin 13, 6-DMAP and KCI is 50.3%, 54.9% and 79.8% respectively. In the KCI treatment of pregnant crabs with the fertilized eggs, the highest induction rate in zoea is 27.3%. Through this study such difficulty as in vitro culture is overcome. Triploid zoea Etiocheir sinensis has been developed for the first time. The induction rate of tetraploid zoea has also been greatly improved.

Zoospores of Undaria pinnatifida: their efficiency to attach under different water velocities and conjugation behavior during attachment

In the invading course of Undaria pinnatifida, zoospore attachment in a dynamically changed subtidal water environment is crucial for the establishment of a potential population in alien waters. Among many abiotic factors that may interfere with the attachment process, water velocity is the most important one. In this investigation, the effect of water velocity on zoospore attachment of U. pinnatifida was investigated in an artificially designed system. It was found that freshly released zoospores that were transported by water flowing at 0 similar to 16 cm/s showed no difficulty in attaching the smooth surface. Zoospore attachment decreased at elevated water flowing rates. At 70 cm/s no spore attachment occurred. Spores that have settled on glass slide for up to I h could not be stripped away by flowing water at a rate of 129 cm/s, the same was true of the 20 d old filamentous gametophytes. It was found that more than 70% of free-swimming zoospores tended to settle down adjacent to the settled spores and formed conjugated clusters from two up to a few hundred cells in still culture.

Population genetics of Crassostrea ariakensis in Asia inferred from microsatellite markers

Crassostrea ariakensis is an important aquacultured oyster species in Asia, its native region. During the past decade, consideration was given to introducing C. ariakensis into Chesapeake Bay, in the United States, to help revive the declining native oyster industry and bolster the local ecosystem. Little is known about the ecology and biology of this species in Asia due to confusion with nomenclature and difficulty in accurately identifying the species of wild populations in their natural environment. Even less research has been done on the population genetics of native populations of C. ariakensis in Asia. We examined the magnitude and pattern of genetic differentiation among 10 wild populations of C. ariakensis from its confirmed distribution range using eight polymorphic microsatellite markers. Results showed a small but significant global theta (ST) (0.018), indicating genetic heterogeneity among populations. Eight genetically distinct populations were further distinguished based on population pairwise theta (ST) comparisons, including one in Japan, four in China, and three populations along the coast of South Korea. A significant positive association was detected between genetic and geographic distances among populations, suggesting a genetic pattern of isolation by distance. This research represents a novel observation on wild genetic population structuring in a coastal bivalve species along the coast of the northwest Pacific.

Distribution of the nonlinear random ocean wave period

Because of the intrinsic difficulty in determining distributions for wave periods, previous studies on wave period distribution models have not taken nonlinearity into account and have not performed well in terms of describing and statistically analyzing the probability density distribution of ocean waves. In this study, a statistical model of random waves is developed using Stokes wave theory of water wave dynamics. In addition, a new nonlinear probability distribution function for the wave period is presented with the parameters of spectral density width and nonlinear wave steepness, which is more reasonable as a physical mechanism. The magnitude of wave steepness determines the intensity of the nonlinear effect, while the spectral width only changes the energy distribution. The wave steepness is found to be an important parameter in terms of not only dynamics but also statistics. The value of wave steepness reflects the degree that the wave period distribution skews from the Cauchy distribution, and it also describes the variation in the distribution function, which resembles that of the wave surface elevation distribution and wave height distribution. We found that the distribution curves skew leftward and upward as the wave steepness increases. The wave period observations for the SZFII-1 buoy, made off the coast of Weihai (37A degrees 27.6' N, 122A degrees 15.1' E), China, are used to verify the new distribution. The coefficient of the correlation between the new distribution and the buoy data at different spectral widths (nu=0.3-0.5) is within the range of 0.968 6 to 0.991 7. In addition, the Longuet-Higgins (1975) and Sun (1988) distributions and the new distribution presented in this work are compared. The validations and comparisons indicate that the new nonlinear probability density distribution fits the buoy measurements better than the Longuet-Higgins and Sun distributions do. We believe that adoption of the new wave period distribution would improve traditional statistical wave theory.

模糊免疫非线性PID控制的优化设计

在深入分析生物免疫系统中T细胞对B细胞辅助调节作用的基础上,提出了免疫反馈原理。针对非最小相位极点系统控制的难点,借鉴免疫反馈原理,结合积分控制的规律,提出了一种模糊免疫非线性PID控制方法。由于该方法中的参数确定比较复杂,利用免疫进化算法进行参数优化设计,实现了控制参数的合理设计。仿真结果表明,该方法在非最小相位极点系统控制中可行且有效,优于PID控制方法,具有更好的响应特性和抗干扰性能。

WIA网络与MODBUS网络互联设计与实现

本文首先介绍了WIA技术和Modbus协议的基本原理和结构,然后详细介绍了互联过程中使用的两种方式:透明传输方式和协议转换方式。分析了两种方式的适用范围和改造难度,并给出了实现案例。

基于UD分解的自适应扩展集员估计方法

用于非线性椭球估计的扩展集员算法在实际应用中存存着数值稳定性差、计算复杂度高以及滤波器参数难以选择等问题.本文提出了一种基于UD分解的自适应扩展集员估计算法,用于解决非线性系统时变状态和参数的联合估计和定界问题.新算法将UD分解与序列更新和选择更新策略结合起米,改进了传统扩展集员算法的数值稳定性和实时性能;同时,对滤波器参数进行自适应选择以进一步降低计算复杂度并达到次优估计结果.仿真实验表明了该算法的有效性和鲁棒性.

基于二维位置敏感探测器的空间对接过程研究

为了准确地捕捉机器人空间对接的传感器信息,从对接机构的设计入手,研究了空间对接中传感器的选择、改制和基于传感器信息的空间对接问题.考虑到安装空间的限制和处理的信息量太多,提出了利用二维位置敏感探测器(PSD)配合红外发光器阵列的方法完成机器人的空间对接.通过对位置敏感探测器工作原理的分析和对传感器特性的试验,提出了在PSD光敏面上增加光学装置的方法以扩大它的探测范围和精度,同时大大减小了它对光信号的依赖程度.把该方法应用于可重构模块化探测机器人系统中,提出了基于传感器信息的轨迹规划方法,对子机器人空间对接过程的仿真试验验证了轨迹规划和空间对接的可行性.

人脸检测的一种混合方法研究

人脸检测作为自动人脸识别系统的第一个环节具有非常重要的作用,为了解决目前大部分人脸检测方法存在的分类器训练困难和检测计算量大等问题,提出了一种人脸检测的混合方法。该方法由两级分类器组成,第一级为粗分类器主要过滤大部分非人脸区域,第二级为核心分类器,在由第一级粗分类的基础上利用非线性SVM算法进行人脸检测。在CMU数据库上的实验结果表明,该方法具有较高的人脸检测率,检测速度得到大幅提高。