大豆种子低温吸胀相关基因的分离与蛋白变化的研究

大豆 (Glycine max (L.) Meer.)是人们日常生活中不可缺少的食品，也是一种非常重要的油质、蛋白资源。目前根据大豆种子吸胀阶段对低温敏感性的不同，可将其划分成3种生态型：低温非敏感型、低温敏感型及中间型。对于低温非敏感型的种子来讲，4℃下吸胀24小时对其发芽率影响很小，而敏感型种子萌发率不超过5%。我国属于温带大陆性气候，大豆播种后由于温度波动而造成一部分种子不能萌发，最终导致减产甚至绝产的现象普遍存在。高产是育种工作的主要目标，提高逆境胁迫的适应能力是高产的前提和基础，所以从分子角度研究种子吸胀非常必要，一方面能够挖掘新的基因资源，另一方面为今后育种工作提供必要的理论依据。 本试验以此为立足点，低温吸胀非敏感型大豆品种 (Z22)为材料，利用cDNA-AFLP方法及蛋白质技术分离与低温吸胀相关的基因及蛋白，得到结果如下： 第一，本试验成功的分离出4个受低温诱导的基因，半定量RT-PCR方法进一步验证了这4个基因在种子吸胀24小时内受低温诱导。 第二，利用RACE方法成功的得到2个完整的全长基因，在NCBI数据库中查找后发现其中1个基因为新基因，命名为SCHI基因 (SCHI：Soybean chilling-induced gene)。SCHI全长为390bp，编码分子量大约为14.2KD的蛋白；另外一个基因是已知基因，其同源序列已经在其他的物种中得到分离。由于此基因与核糖体蛋白L34高度同源，所以把把这个基因命名为SOL34 (Soybean L34)。 第三，利用半定量RT-PCR方法对基因表达模型进行分析，结果表明：SCHI在种子低温吸胀18～24小时期间诱导表达量最高，而当种子低温吸胀24小时后转入常温下，其表达量在常温下18小时左右迅速下降；ABA (100μM)、PEG (30%，10000)及NaCl (250mM)能够诱导SCHI的表达，在诱导表达量上，ABA和PEG诱导效果最明显，而NaCl能够微弱的诱导此基因表达；对不同生态型的大豆品种而言，低温吸胀过程中，SCHI在非敏感型种子中的表达量高于敏感型种子，但非敏感型和中间型之间没有差别；另外，SCHI在大豆胚轴中是诱导型表达，在叶片和根尖中则是组成型表达。SOL34的表达在萌发前24小时内被低温诱导，但在不同生态型之间没有差别。SOL34在胚轴和根尖中受低温诱导，在叶片中是组成型表达。 第四，SCHI能够在原核生物中表达出相应蛋白，诱导表达蛋白的分子量在26-29KD，大约为理论值的2倍，说明在大肠杆菌中被表达的蛋白以2聚体形式存在。另外低温试验结果表明SCHI能够提高菌落忍耐短时间-20℃低温的能力。 第五，利用双元表达载体把SCHI转入拟南芥植株，经过低温、干旱和盐胁迫后，转基因植株的成活率均高于野生型植株。超表达SOL34的拟南芥植株降低了对低温的耐性；而抑制拟南芥中L34的表达反而提高了植株对低温的抗性。 第六，本试验利用蛋白质等有关试验检测了大豆种子低温吸胀时蛋白质发生的变化。从吸胀 (4℃和22℃下24h)后的大豆胚轴中成功鉴定出上调蛋白点25个，下调蛋白点15个。其中有参与能量代谢反应 (占10%，例如柠檬酸脱氢酶和苹果酸脱氢酶等)、细胞生长与分裂相关反应 (20%，例如LEA蛋白和种子成熟蛋白PM26)、胁迫反应 (10%，如乙醇脱氢酶)、种子宿命和贮藏蛋白 (20%，大豆球蛋白)等蛋白在此过程中发生了变化，暗示种子萌发前期低温吸胀过程中多种代谢发生变化。细胞生长变缓、能量代谢增强、胁迫代谢蛋白的高表达以及贮藏蛋白降解速度减慢等变化都有利于种子在吸胀过程中度过低温环境，为以后的生长作好准备。

Investigating the relationships between user capabilities and product demands for older and disabled users

This paper presents the results of a study that specifically looks at the relationships between measured user capabilities and product demands in a sample of older and disabled users. An empirical study was conducted with 19 users performing tasks with four consumer products (a clock-radio, a mobile phone, a blender and a vacuum cleaner). The sensory, cognitive and motor capabilities of each user were measured using objective capability tests. The study yielded a rich dataset comprising capability measures, product demands, outcome measures (task times and errors), and subjective ratings of difficulty. Scatter plots were produced showing quantified product demands on user capabilities, together with subjective ratings of difficulty. The results are analysed in terms of the strength of correlations observed taking into account the limitations of the study sample. Directions for future research are also outlined. © 2011 Springer-Verlag.

Modelling cognitive impairment to improve universal access

The purpose of this study is to develop a model of cognitive impairment to help designers consider the range of issues which affect the lives of people living with such impairment. A series of interviews with experts of cognitive impairment was conducted to describe and assess the links between specific medical conditions, including learning disability, specific learning difficulties, autistic spectrum disorders, traumatic brain injury and schizophrenia, and the types of cognitive impairment associated with them. The results reveal some of the most prevalent and serious types of impairment, which - when transformed into design guidance - will help designers make mainstream products more inclusive also for people with cognitive impairment. © 2011 Springer-Verlag.

User responses to the learning demands of consumer products

Designers are typically male, under 35 years old and unimpaired. Users can be of any age and currently over 15% will have some form of impairment. As a result a vast array of consumer products suit youthful males and in many cases exclude other demographics (e.g. Keates and Clarkson, 2004). In studying the way a range of users learn how to use new products, key cognitive difficulties are revealed and linked back to the areas of the product causing the problems. The trials were structured so each user had to complete a specific set of tasks and were consistent across the user spectrum. The tasks set aimed to represent both everyday usage and less familiar functions. Whilst the knowledge gained could provide designers with valuable guidelines for the specific products examined, a more general abstraction provides knowledge of the pitfalls to avoid in the design of other product families.

Designing inclusive interfaces through user modeling and simulation

Elderly and disabled people can be hugely benefited through the advancement of modern electronic devices, as those can help them to engage more fully with the world. However, existing design practices often isolate elderly or disabled users by considering them as users with special needs. This article presents a simulator that can reflect problems faced by elderly and disabled users while they use computer, television, and similar electronic devices. The simulator embodies both the internal state of an application and the perceptual, cognitive, and motor processes of its user. It can help interface designers to understand, visualize, and measure the effect of impairment on interaction with an interface. Initially a brief survey of different user modeling techniques is presented, and then the existing models are classified into different categories. In the context of existing modeling approaches the work on user modeling is presented for people with a wide range of abilities. A few applications of the simulator, which shows the predictions are accurate enough to make design choices and point out the implication and limitations of the work, are also discussed. © 2012 Copyright Taylor and Francis Group, LLC.

Universal access and assistive technology

This book contains the proceedings of the first Cambridge Workshop on Universal Access and Assistive Technology (CWUAAT), incorporating the fourth Cambridge Workshop on Rehabilitation Robotics, held in Cambridge, England in March 2002.

Designing Accessible Technology

The key to this is maintaining a healthy population that is able and willing to work longer before retirement and can remain independent for as long as possible afterwards as well as bringing disabled people into mainstream life and ...

Designing inclusive systems

The Cambridge Workshops on Universal Access and Assistive Technology (CWUAAT) are a series of workshops held at a Cambridge University College every two years.

A population perspective on mobile phone related tasks

Chapter 6 A Population Perspective on Mobile Phone Related Tasks M. Bradley, S. Waller, J. Goodman-Deane, l. Hosking, R. Tenneti, PM Langdon and PJ Clarkson 6.1 Introduction For design to be truly inclusive, it needs to take into ...