鲸类分子系统学研究进展

综述了近年来分子生物学标记技术在鲸类系统学研究中的进展。分子生物学证据支持鲸目与有蹄类之间有较近的亲缘关系,并支持鲸类的单系起源,但鲸类不同类群(须鲸类、抹香鲸类及不包括抹香鲸类的齿鲸类)之间的系统发生关系仍存在争议。抹香鲸类到底与须鲸类还是与其它齿鲸类有更近的亲缘关系,不同的分子生物学家所得到的结果并不一致。此外,分子生物学技术还被用于解决须鲸亚目和齿鲸亚目内科间以及科内种间的系统发生关系,特别是齿鲸亚目的海豚科、鼠豚科和淡水豚类。通过分子标记技术来研究鲸类种下的遗传结构是鲸类分子系统学研究中的一个新热点,使用的标记主要是mtDNA控制区、核DNA微卫星和主要组织相容性复合体(major histo-compatibilitv complex,MHC)等。     

兰花组织培养和分子生物学研究进展

兰科植物是开花植物中最大的家族之一,其科研和经济价值越来越受到全世界的重视。兰花的组织培养近年来发展迅速,对兰花组织培养中原球茎的诱导和培养基选择的国内外研究进行了综述;并对近年来应用分子标记、转基因等分子生物学技术研究兰花的遗传多样性、系统分类和基因功能进行综述。     

Doubled haploid production in Flax (Linum usitatissimum L.)

There is a requirement of haploid and double haploid material and homozygous lines for cell culture studies and breeding in flax. Anther culture is currently the most successful method producing doubled haploid lines in flax. Recently, ovary culture was also described as a good source of doubled haploids. In this review we focus on tissue and plants regeneration using anther culture, and cultivation of ovaries containing unfertilized ovules. The effect of genotype, physiological status of donor plants, donor material pre-treatment and cultivation conditions for flax anthers and ovaries is discussed here. The process of plant regeneration from anther and ovary derived calli is also in the focus of this review. Attention is paid to the ploidy level of regenerated tissue and to the use of molecular markers for determining of gametic origin of flax plants derived from anther and ovary cultures. Finally, some future prospects on the use of doubled haploids in flax biotechnology are outlined here.     

Molecular genetic tools to infer the origin of forest plants and wood

Most forest tree species exhibit high levels of genetic diversity that can be used to trace the origin of living plants or their products such as timber and processed wood. Recent progress to isolate DNA not only from living tissue but also from wood and wood products offers new opportunities to test the declared origin of material such as seedlings for plantation establishment or timber. However, since most forest tree populations are weakly differentiated, the identification of genetic markers to differentiate among spatially isolated populations is often difficult and time consuming. Two important fields of “forensic” applications are described: Molecular tools are applied to test the declared origin of forest reproductive material used for plantation establishment and of internationally traded timber and wood products. These applications are illustrated taking examples from Germany, where mechanisms have been developed to improve the control of the trade with forest seeds and seedlings, and from the trade with wood of the important Southeast Asian tree family Dipterocarpaceae. Prospects and limitations of the use of molecular genetic methods to conclude on the origin of forest plants, wood, and wood products are discussed.     

In vitro production of haploid and doubled haploid plants from pollinated ovaries of maize (Zea mays)

Haploid induction has potential application for maize breeding. This paper reports that maize haploid plants have been induced by in vitro culture of pollinated ovaries. From a total of 26,400 cultured ovaries, 24 haploid plants were obtained and two of them were doubled after colchicine treatment. The maximum frequency of gynogenesis was 0.17% at 19.5 h post-pollination (HPP). The results showed that HPP was an important factor affecting plant induction from ovaries. Regenerated diploid R₀ plants were then subjected to genetic analysis using SSR molecular markers. One R₀ plant, whose progeny revealed a high level of homogeneity for several agro-morphological traits, was homozygous at 20 loci tested, with 11 showing paternal and 9 maternal banding pattern. This demonstrates that it is feasible to induce maize haploid plants by in vitro culture of pollinated ovaries.     

Molecules and migration: biogeographical studies in cruciferous plants

In the past two decades our understanding of plant biogeography has been improved substantially by the introduction of various molecular marker systems. Especially within the angiosperms, maternally inherited chloroplast DNA based data sets have elucidated not only genetic relatedness but also geographic structuring of genetic variation. These findings were based on the observation that DNA molecules might mutate during migration, which consequently found its manifestation in the term phylogeography introduced in the late 80s by John Avise. However, other markers such as codominantly inherited allozymes were used before the advent of DNA techniques and were used in theoretical population genetic studies. In actual phylogeographic studies, highly variable markers, such as AFLPs (amplified fragment length polymorphisms), were needed to unravel recent species histories (e.g. pleistocenic differentiation). The levels of molecular variation at such markers are closer to that of allelic variation measured with allozymes. Hence, an increasing number of studies have relied on highly polypmorphic markers, such as DNA microsatellite loci. Herein, we try to present an overview on the various biogeographic and phylogeographic studies using various molecular (including isozyme) markers and methodological approaches to analyse them, concentrating on studies done with representatives of the Brassicaceae family.     

Improving salinity tolerance in crop plants: a biotechnological view

Salinity limits the production capabilities of agricultural soils in large areas of the world. Both breeding and screening germplasm for salt tolerance encounter the following limitations: (a) different phenotypic responses of plants at different growth stages, (b) different physiological mechanisms, (c) complicated genotype × environment interactions, and (d) variability of the salt-affected field in its chemical and physical soil composition. Plant molecular and physiological traits provide the bases for efficient germplasm screening procedures through traditional breeding, molecular breeding, and transgenic approaches. However, the quantitative nature of salinity stress tolerance and the problems associated with developing appropriate and replicable testing environments make it difficult to distinguish salt-tolerant lines from sensitive lines. In order to develop more efficient screening procedures for germplasm evaluation and improvement of salt tolerance, implementation of a rapid and reliable screening procedure is essential. Field selection for salinity tolerance is a laborious task; therefore, plant breeders are seeking reliable ways to assess the salt tolerance of plant germplasm. Salt tolerance in several plant species may operate at the cellular level, and glycophytes are believed to have special cellular mechanisms for salt tolerance. Ion exclusion, ion sequestration, osmotic adjustment, macromolecule protection, and membrane transport system adaptation to saline environments are important strategies that may confer salt tolerance to plants. Cell and tissue culture techniques have been used to obtain salt tolerant plants employing two in vitro culture approaches. The first approach is selection of mutant cell lines from cultured cells and plant regeneration from such cells (somaclones). In vitro screening of plant germplasm for salt tolerance is the second approach, and a successful employment of this method in durum wheat is presented here. Doubled haploid lines derived from pollen culture of F₁ hybrids of salt-tolerant parents are promising tools to further improve salt tolerance of plant cultivars. Enhancement of resistance against both hyper-osmotic stress and ion toxicity may also be achieved via molecular breeding of salt-tolerant plants using either molecular markers or genetic engineering.     

Germplasm-regression-combined (GRC) marker-trait association identification in plant breeding: a challenge for plant biotechnological breeding under soil water deficit conditions

In the past 20 years, the major effort in plant breeding has changed from quantitative to molecular genetics with emphasis on quantitative trait loci (QTL) identification and marker assisted selection (MAS). However, results have been modest. This has been due to several factors including absence of tight linkage QTL, non-availability of mapping populations, and substantial time needed to develop such populations. To overcome these limitations, and as an alternative to planned populations, molecular marker–trait associations have been identified by the combination between germplasm and the regression technique. In the present preview, the authors (1) survey the successful applications of germplasm–regression–combined (GRC) molecular marker–trait association identification in plants; (2) describe how to do the GRC analysis and its differences from mapping QTL based on a linkage map reconstructed from the planned populations; (3) consider the factors that affect the GRC association identification, including selections of optimal germplasm and molecular markers and testing of identification efficiency of markers associated with traits; and (4) finally discuss the future prospects of GRC marker–trait association analysis used in plant MAS/QTL breeding programs, especially in long-juvenile woody plants when no other genetic information such as linkage maps and QTL are available.     

Biotechnological advances in pomegranate (Punica granatum L.)

Pomegranate (Punica granatum L.) is known for its nutritional, medicinal, and ornamental importance. It is conventionally propagated by hardwood and softwood cuttings, but about 1 yr is needed before the rooted cuttings can be transplanted to the field. Propagation by seed is undesirable as populations are heterozygous and seed propagation leads to wide variations in tree and fruit characteristics. Several studies have been conducted on in vitro culture of pomegranate, and protocols have been developed for plant regeneration through organogenesis and embryogenesis from various types of explants. Tissue culture has enabled mass propagation of superior genotypes of both wild and cultivated varieties. However, successful application of tissue culture systems for genetic engineering of pomegranate is still limited. Molecular markers are essential for identification and discrimination of genotypes for genetic conservation, crop improvement, breeding programs, and commercialization of superior genotypes. These techniques may also be applicable to rapid identification and indexing of disease-free planting material. This review focuses on the biotechnological approaches that are being used for pomegranate improvement.     

分子标记在濒危物种保护中的应用

分子标记可揭示种群遗传和进化信息, 为制定濒危物种保护措施、指导恢复实践提供重要依据。本文主要介绍了分子标记在濒危物种保护过程不同环节中的应用, 包括: (1)正确识别保护单元, 如排除隐存种和杂交种的影响; (2)确定优先保护单元, 包括优先保护区域、优先保护物种、优先保护种群等; (3)指导迁地保护; (4)对保护工作的动态监测和评估。文章最后探讨了分子标记应用于保护的发展方向, 如开展长期的种群遗传组成监测、切实应用于保护管理实践、将基因组学等遗传信息用于全球变化背景下保护策略的制定等, 期望为分子标记技术在生物多样性保护的研究和实践中提供参考。     

Plant tissue culture. Biotechnology. Milestones

Summary The progress in the development of the technologies of plant tissue and cell culture over the past four decades has been remarkable. This article covers my personal reflections on the various topics and is based on my involvement in the field during that period. There are three fundamental technologies which constitute most of what is referred to as plant in vitro technologies or tissue culture. The origin and some of the key persons involved in the development of each of these procedures will be discussed. The technology that is most common is growing plant tissue on gel-solidified nutrient media. That technology is being used in the most vital procedures, namely the regeneration of plants from cultured cells. The culture of plant cells in liquid suspension was developed very shortly after that, and has become a very effective technology for plant regeneration by somatic embryogenesis. The method of meristem culture arose out of a need for developing plants that were virus-free. In many species the technique is now being used to produce virus-free crop plants. Another important technology is the culture of anthers and microspores for producing haploid and homozygous plants. Included with plant tissue culture is the development of the plant protoplast and cell fusion technologies for the production of new plant hybrids. The final aspect of the development concerns the integration of tissue culture with molecular genetics, which has developed into the rapidly expanding field of biotechnology.     

白蚁的分子生物学研究进展

白蚁是危险性社会昆虫,建立安全有效的白蚁防治方法有赖多学科的参与,分子生物学已经成为白蚁研究的重要工具。目前,DNA序列分析方法已应用于白蚁鉴定和分类,其中线粒体基因是最通用的分子标记;基因工程技术成功构建了可用于白蚁防治的白蚁肠道工程菌;Hexamerin、COX Ⅲ、纤维素酶等白蚁功能基因以及白蚁品级分化相关的若干蛋白相继得到了分离和鉴定。文章从白蚁分类、防治、功能基因、品级分化4个方面综述白蚁分子生物学的研究进展,为白蚁的防治提供新的方法和思路。     

基于DNA分子标记的花粉流动态分析

花粉介导的基因流是植物有性繁殖世代之间的桥梁, 花粉散布属性是植物繁殖生态学、保护生物学和进化生物学研究关注的焦点。随着DNA分子技术的发展, 花粉流分析所使用的分子标记(尤其是微卫星标记)逐步替代了早期物理标记, 基于最大似然法估计以及新兴的基于贝叶斯推断的父本指派算法的发展, 能有效地估计花粉流散布的方向、距离和强度等重要特征。花粉散布曲线由单一参数向多参数模型发展, 以更好地获得花粉散布特征的拟合效果, 双组分的复合模型利用相互独立的参数空间使得散布曲线在长距离和短距离形状上呈现更大的可塑性。这些革新的技术和方法被成功应用于植物性别表型、隔离种群和杂交物种间花粉流分析, 以探讨进化、生态和保护等多领域的基础理论问题。近年来, 高通量测序技术的发展将进一步加快以分子标记为基础的花粉流动态分析在更广泛的植物类群中运用。     

Camptothecin from <Emphasis Type="Italic">Nothapodytes nimmoniana</Emphasis>: review on biotechnology applications

Since the first report on Camptothecin detection in Nothapodytes nimmoniana by Govindachari and Viswanathan (Phytochem 11:35–29, 1972), considerable work has been done on biotechnology and its applications on the species. Plant tissue culture techniques have applications in clonal propagation, CPT production, and conservation of N. nimmoniana. Discovery of CPT production by endophytes existing in symbiotic association with N. nimmoniana has provided new insights into finding alternative sources of the alkaloid. Development of molecular markers such as RFLP, RAPD, ISSR, and AFLP has facilitated understanding of population ecology and genetics of the species. Molecular information generated from these studies is promising in establishing strategies for conservation and sustainable use of N. nimmoniana populations under overexploitation pressure. The advances in instrumentation in the 20th century, such as desorption electrospray ionization mass spectrometry allowed CPT analysis in tissues without sample pretreatment. Other ancient techniques for qualitative and quantitative analysis such as chromatography, spectroscopy, and H1-NMR are applied in the detection of CPT due to variable sensitivity to the alkaloid. The review covers work on plant tissue culture for clonal propagation and CPT production in N. nimmoniana. Besides symbiotic endophyte sources of CPT in N. nimmoniana, population genetics studies and instrumentation analysis of the alkaloids are reviewed.     

热带与亚热带森林分子生态学研究

当前 ,植物学家会有更多的方法和手段来从事植物学的研究 ,如分子生物学方法、遗传学的方法、植物生理学的方法、分类学方法等。其中分子生物学方法直接研究植物遗传物质ＤＮＡ ,从根本上揭示了生物体内涵 ,有着极其重要的作用[4 7] 。因此用分子生物学手段进行生态学的研究有着广泛应用前景和重要意义。分子生物学方法既节省时间和资金 ,又比传统方法解决了更多的问题[6 1] 。例如在动物学研究中 ,用少许的毛发就可以分析高山土拨鼠 (ＭａｒｍｏｔａｍａｒｍｏｔａＳｃｉｕｒｉｄａｅ)的交配系统[30 ] ;粪便所带的肠道粘膜细胞ＤＮＡ也可以…     

Genetic diversity analysis of 60 ginger germplasm core accessions using ISSR and SSR markers

Molecular techniques play a critical role in studies of phylogeny and, thus, have been applied to understand the distribution and extent of genetic variation within and between species. In the present study, a genetic analysis was undertaken using molecular markers (9 ISSR and 13 SSR) on 60 ginger cultivars from different regions of the eastern coast of India (Odisha). The data obtained with 22 polymorphic markers revealed moderate to high diversity in the collection. Both ISSR and SSR markers were efficient in distinguishing all the 60 ginger cultivars. A total of 42 and 160 polymorphic bands were observed with ISSR and SSR markers, respectively. However, SSR markers were observed to be better at displaying average polymorphism (63.29%) than ISSR markers (55%). Analysis of molecular variance results showed that 52 and 66% of the variation occurred among different ginger populations, whereas 48 and 34% of the variation was found within populations, respectively, using ISSR and SSR markers, indicating that ginger cultivars display significant genetic diversity at the population level. Principal coordinates analysis and the dendrogram constructed out of combined data of both markers showed grouping of ginger accessions to their respective area of collection, indicating geographical closeness due to genetic similarity irrespective of the relationship that exists at the morphological level.     

Isolated microspore culture techniques and recent progress for haploid and doubled haploid plant production

An isolated microspore culture provides an excellent system for the study of microspore induction and embryogenesis, provides a platform for an ever-increasing array of molecular studies, and can produce doubled haploid (DH) plants, which are used to accelerate plant-breeding programs. Moreover, isolated microspore cultures have several advantages over anther culture, wherein presence of the anther walls can lead to the development of diploid, somatic calli and plants. Although protocols for isolated microspore culture vary from laboratory to laboratory, the basic steps of growing donor plants, harvesting floral organs, isolating microspores, culturing and inducing microspores, regenerating embryos, and doubling the chromosomes, remain the same. Over the past few years, a large proportion of the research reports on isolated microspore culture have focused on cereal and Brassica species. For some of these species, isolated microspore culture protocols are well established and routinely used in laboratories around the world for developing new varieties, as well as for basic research in areas such as genomics, gene expression, and genetic mapping. Although these species are considered highly responsive to microspore culture, improvements in efficiency are still being made. However, with many species, isolated microspore culture is simply not yet efficient enough at producing DH plants to be cost-effective for breeding programs. There has been a recent resurgence of haploidy research with response being reported in some species once considered recalcitrant. Future research programs aimed at elucidating pathways involved in microspore induction and embryogenesis will be of benefit, as will novel approaches to improve the efficiency of microspore culture for DH production. With many species, anther culture has proven to be more effective than isolated microspore culture, necessitating more research to clarify the contribution of the anther wall to embryogenesis. The development of molecular markers for use in determining the gametic origin of regenerated plants, irrespective of their ploidy, would also be beneficial. In this review, we aim to provide an overview of the basic isolated microspore culture protocol with an emphasis on recent progress in several crop species.     

Anther culture in pepper (<Emphasis Type="Italic">Capsicum annuum</Emphasis> L.) in vitro

Pepper (Capsicum annuum L.) is an important vegetable crop that can be improved using plant tissue culture and biotechnology. However, it is difficult to develop appropriate breeding material by in vitro cultivation in this species. Haploid plant production is useful in the breeding programs to facilitate recovery of recessive mutations and unique genetic recombinations. In embryogenesis, haploid formation from pollen in anther culture is a scientifically advanced, but controversial system. Various techniques for haploid plant regeneration are used to establish an efficient double haploid production method. The purpose of this article is to summarize, through comparison, results in pepper anther culture, problems associated with work in this field, and the influence of critical factors for successful embryo formation and plantlet development.