裸燕麦EMS突变体库筛选与分析

燕麦是重要的粮饲兼用作物,构建燕麦EMS突变体库对燕麦功能基因组学研究和遗传改良有重要意义。本试验利用化学诱变剂甲基磺酸乙酯(EMS,ethyl methane sulfonate)处理燕麦品种花早2号,获得了4083株M1材料;对其中2000个单株种植了M2株行,进行全生育期调查,鉴定其表型变化;对2份黄化苗突变材料种植了M3家系,进行相关突变性状的稳定性验证。结果表明,燕麦经EMS处理后代变异巨大,在M2发现表型突变材料196份,变异率为9.8%,变异类型非常丰富,包括幼苗习性、叶片性状、分蘖、株高、穗部形态及成熟期等突变株系。M3证实突变的黄化苗特性可以稳定遗传。本研究建立了燕麦EMS诱变体系,获得的燕麦变异类型丰富,为燕麦功能基因组学研究和燕麦遗传改良奠定了材料基础。     

TILLING技术及其应用

定向诱导基因组局部突变(targetinginducedlocallesionsingenomes,TILLING)可快速、有效地鉴定和定向筛选突变,是一种全新的反向遗传学技术。现对TILLING的技术流程、核心与特点,及其在突变研究、反向遗传学及功能基因组学、SNP检测、资源创新与分析以及作物遗传改良等方面的应用进行了综述。     

蔬菜作物的组织培养

经济作物中有一些属于杂合子型植物(heterozygote),例如马铃薯、球茎和鳞茎植物以及果树、花卉等植物。这类植物如用种子繁殖,它们的后代便会出现各种变异,不能保持原来品种的特性,因此一般均采用无性繁殖。近年来植物组织和细胞培养的技术与研究获得了飞跃的发展,应用这一技术来繁殖蔬菜作物,不仅使繁殖速度可以极大地增加,而且使不易用常规无性繁殖方法来繁殖的植物,也获得了新的繁殖途     

RNAi技术在作物品质改良中的应用

RNA干涉(RNAi)是由同源性内源或外源dsRNA引起的序列特异性基因沉默现象,在动、植物和真菌中广泛存在并被证实。目前已应用RNAi技术在改善油脂的品质、改良淀粉品质、提高营养物质或降低有害物质含量、提高抗褐化能力、提高果实耐贮性、进行代谢调控以获得目的次生代谢物等方面进行了作物品质改良研究。作为一种下调表达技术,该技术在研究植物基因功能和改良作物品质等领域有良好的应用前景。本文重点从上述六个方面对近年来应用RNAi技术在作物品质改良研究方面进行了回顾并对其存在的问题作了初步探讨。     

法国用诱变育种技术替代转基因农作物

为改良传统农作物的品种和性能,法国采用植物诱发突变技术育种已有近80年的历史。据世界粮农组织估计,2007年全球共种植2550种诱变育种农作物,其中近一半为粮食作物,生态农田也采用诱变育种的农作物品种。诱变育种农作物与转基因农作物不同,主要利用物理或化学的因素处理作物,使其发生基因突变。基因突变是作物变异进化的根本来源,为作物进化提供了最初的原材料。     

转基因改良植物的营养价值

植物是人类所需大部分营养物质的主要来源,植物产品的营养品质直接影响着人类的健康。分子克隆和遗传转化技术的发展为改良植物的营养价值开辟了新途径。植物营养价值的转基因改良已在改进作物蛋白质含量及品质、淀粉和油脂成分及品质,提高抗氧化物水平（如类胡萝卜素、类黄酮等）,培育具有医疗效应的营养品质等方面取得了可喜的进展。迄今,已获得许多营养品质改良的转基因作物品系。这些转基因作物经过一系列的安全性及对人类营养有效性的验证后, 可直接食用,或应用于开发具有特殊营养品质和保健作用的“功能食品”。我们实验室开展了大豆油脂改良研究,构建了能特异抑制大豆FAD2-1基因表达的锌指转录因子,获得了油酸含量显著提高的转基因材料。初步结果表明锌指转录因子的分子设计是改良植物油脂代谢的一条可行途径,亦可用于调控植物其它内源靶基因的表达。     

假单胞杆菌BS1转座突变及高产生物表面活性剂突变株初步筛选

利用含转座子Tn917的温敏性质粒pTV1-OK转化假单胞杆菌BS1原生质体,成功获得3个稳定的转化子;通过Tn917诱导转座突变,产生大量突变体,构建了转座突变体库,采用特异引物对随机挑取的5株突变体进行PCR扩增,获得与预期大小一致片段,表明突变体基因组中有Tn917插入;通过对随机挑取24株突变体乳化(E24)性能测定,发现有一株突变体E24值达到67%,明显高于野生菌株。结果表明转座突变是假单胞杆菌BS1获得高产生物表面活性剂菌株的一种有效手段。     

RNA干扰及其在果树上的应用

RNA干扰是广泛存在于生物中的一种现象,它是由小干扰RNA诱导的特异基因沉默.它是生物体抵抗异常的一种防御机制,同时在生物生长发育过程中调控基因的表达,为植物基因功能的研究开辟了新途径.综述了RNA干扰的作用机制、RNA干扰的特点,以及近年来RNA干扰在果树生长发育、抗病性、果实品质改良等方面的应用,以期为RNA干扰技术在果实品种品质改良中的应用提供指导.     

未来的一种生物肥料:丛枝菌根真菌*

丛枝菌根真菌(Arbuscular Mycorrhizal Fungi,AMF)存在于几乎所有类型的土壤中,可以与绝大多数被子植物的根共生。大多数农作物、果树、蔬菜、观赏植物和花卉等都能形成丛枝菌根。AMF能促进作物吸收利用矿质养分和水分,提高作物抗逆性和抗病性,改良土壤、提高苗木移栽成活率、促进生长、提高产量和改善品质,并且可用于改善退化生态系统的土壤肥力,维持农林业的可持续发展,将成为一种新型的生物肥料被用于农林业生产。本文讨论了影响菌根侵染率的因素、AMF的生态效应和在生态农业中的应用现状和前景。     

离体繁育突变体的低成本方法

虽然已用离体突变繁育法成功地获得了花色变异体,但这一技术形成了大量不稳定,低潜力突变体。大量此类“不适”突变体提高了田间试验的成本。有一种假设认为,通过继代培养辐射过的外植体,能够只保留适宜的突变细胞并使之表达,Cork大学的A.C.Cassells及共同事根据这一假设进行试验,尝试在田间试验之前,通过对辐射过的离体芽进行一系列继代培养,清除不适用的变异体。     

Mutation induction and tissue culture in improving fruits

This review describes in vitro mutation induction methods in fruits and the in vitro selection procedures available for early screening. Results obtained through in vitro mutation techniques, including somaclonal variation, are reviewed and compared with the current achievements and future prospects of transgenic breeding. Plant improvement based on mutations, which change one or a few specific traits of a cultivar, can contribute to fruit improvement without altering the requirements of fruit industry. Induced mutations have well defined limitations in fruit breeding applications, but their possibilities may be expanded by the use of in vitro techniques. Tissue culture increases the efficiency of mutagenic treatments for variation induction, handling of large populations, use of ready selection methods, and rapid cloning of selected variants. Molecular techniques can provide a better understanding of the potential and limitations of mutation breeding e.g. molecular marker-assisted selection, which can lead to the early identification of useful variants. The relatively high number of research reports compared with the low number of cultivars released suggests that mutagenesis in combination with tissue culture is either ineffective or has yet to be exploited in fruits. Positive achievement recorded in other species seem to support the hypothesis that in vitro mutation induction has high potential also for fruit improvement. The possible contribution of a well-pondered and coordinated use of the numerous mutation induction, mutant selection, and field validation procedures available to advances in fruit breeding is discussed. This revised version was published online in June 2006 with corrections to the Cover Date.     

Generation of chemically induced mutations using in vitro propagated shoot tip tissues for genetic improvement of fruit trees

Mutagenesis via treatment of seeds with chemical mutagens such as ethyl methanesulfonate (EMS) has been widely used for crop improvement. However, this approach is not suitable for some crop species such as clonally propagated crops and allogamous perennial plants characteristically with high levels of genome heterozygosity and a long life cycle. Here, we report direct treatment of in vitro-induced peach shoot tip tissues with EMS for generation of mutations in peach, a perennial, woody fruit tree. We optimized EMS dosage and exposure time and found that treatment of the explants with 0.2 % EMS for 16 h was optimal for generation of genetic variations. So far we have generated nearly 2000 peach seedlings. Typical EMS-induced phenotypic variations were present in the seedlings. Next generation sequencing and subsequent data analyses were performed to monitor the efficiency of mutagenesis. The mutation rate was estimated to be one mutation per 150 kilobase pairs in the mutagenized population, suggesting effectiveness of this method in inducing mutagenesis in peach. Taken together, our data open an avenue for the generation of mutant populations suitable for crop improvement in allogamous perennial plants including fruit trees and clonally propagated plants.

     

Mapping of two suppressors of OVATE (sov) loci in tomato

Tomato fruit shape varies significantly in the cultivated germplasm. To a large extent, this variation can be explained by four genes including OVATE. While most varieties with the OVATE mutation bear elongated fruits, some accessions carry round fruit, suggesting the existence of suppressors of OVATE in the germplasm. We developed three intraspecific F₂ populations with parents that carried the OVATE mutation but differed in fruit shape. We used a bulk segregant analysis approach and genotyped the extreme classes using a high-throughput genotyping platform, the SolCAP Infinium Assay. The analyses revealed segregation at two quantitative trait loci (QTLs), sov1 and sov2. These loci were confirmed by genotyping and QTL analyses of the entire population. More precise location of those loci using progeny testing confirmed that sov1 on chromosome 10 controlled obovoid and elongated shape, whereas sov2 on chromosome 11 controlled mainly elongated fruit shape. Both loci were located in intervals of <2.4 Mb on their respective chromosomes.     

Establishment of a dmp based maternal haploid induction system for polyploid Brassica napus and Nicotiana tabacum

Doubled haploid (DH) technology is used to obtain homozygous lines in a single generation, a technique that significantly accelerates the crop breeding trajectory. Traditionally, in vitro culture is used to generate DHs, but this technique is limited by species and genotype recalcitrance. In vivo haploid induction (HI) through seed is widely and efficiently used in maize and was recently extended to several other crops. Here we show that in vivo HI can be triggered by mutation of DMP maternal haploid inducer genes in allopolyploid (allotetraploid) Brassica napus and Nicotiana tabacum. We developed a pipeline for selection of DMP orthologs for clustered regularly interspaced palindromic repeats mutagenesis and demonstrated average amphihaploid induction rates of 2.4% and 1.2% in multiple B. napus and N. tabacum genotypes, respectively. These results further confirmed the HI ability of DMP gene in polyploid dicot crops. The DMP-HI system offers a novel DH technology to facilitate breeding in these crops. The success of this approach and the conservation of DMP genes in dicots suggest the broad applicability of this technique in other dicot crops.     

番茄果实早期发育的分子生理机制研究进展

番茄(Solanum lycopersicum)是目前世界上种植面积最广且最受欢迎的蔬菜作物之一, 也是肉果及茄科的重要模式植物。番茄果实发育主要分为早期果实发育和果实成熟2个时期, 但果实形态结构和大小主要决定于早期果实发育时期。该文围绕番茄早期果实发育时期植物激素、细胞周期、表观遗传和源库代谢等多方面调控的分子机制进行了综述, 旨在认识植物生长与发育的基本生物学问题及促进基础理论研究成果在生产中应用。     

Phenological growth stages of jackfruit (Artocarpus heterophyllus) according to the extended BBCH scale

Jackfruit, the largest known edible fruit bearing tree, is one of the important fruit crops of India. It exhibits wide range of diversity with respect to fruit character and bulb quality. It is an important component of homestead garden because of its multifarious uses (dessert, vegetable and pickle) and high nutritive value. In spite of high food value and market potential, jackfruit is one of the poorly researched crops and there is a dearth of information about its phenology. The present study defines phenological stages of jackfruit according to the extended BBCH (Biologische Bundesantalt, Bundessortenamt und Chemische Industrie) scale using three‐digit numerical system. Eight principal growth stages, namely bud development (stage 0), shoot development (stage 1), leaf development (stage 3), specialised reproductive shoot development (stage 4), reproductive development (stage 5), flowering (stage 6), fruit development (stage 7) and fruit maturation (stage 8) have been described. A total of 42 secondary growth stages have been described and defined. In this study, bearing and non‐bearing footstalks as well as male and female phases have been defined separately. The study will act as an effective tool for providing a consensual unified approach for standardisation of phenophases, as well as for efficient orchard management for ensuring higher yield and fruit quality. The scale may also be effectively used for characterisation and adaptation of germplasm and assessment of climatic impact on crop phenology.     

Papaya Ring Spot Virus: An Understanding of a Severe Positive-Sense Single Stranded RNA Viral Disease and Its Management

Viral diseases have been studied in-depth for reducing quality, yield, health and longevity of the fruit, to highlight the economic losses. Positive-sense single-stranded RNA viruses are more devastating among all viruses that infect fruit trees. One of the best examples is papaya ringspot virus (PRSV). It belongs to the genus Potyvirus and it is limited to cause diseases on the family Chenopodiaceae, Cucurbitaceae and Caricaceae. This virus has a serious threat to the production of papaya, which is famous for its high nutritional and pharmaceutical values. The plant parts such as leaves, latex, seeds, fruits, bark, peel and roots may contain the biological compound that can be isolated and used in pharmaceutical industries as a disease control. Viral disease symptoms consist of vein clearing and yellowing of young leaves. Distinctive ring spot patterns with concentric rings and spots on fruit reduce its quality and taste. The virus has two major strains P and W. The former cause disease in papaya while the later one in papaya. Virion comprises 94.4% protein, including a 36 kDa coat protein which is a component responsible for a non-persistent transmission through aphids, and 5.5% nucleic acid. Cross protection, development of transgenic crops, exploring the resistant sources and induction of pathogen derived resistance have been recorded as effective management of PRSV. Along with these practices reduced aphid population through insecticides and plant extracts have been found ecofriendly approaches to minimize the disease incidence. Adoption of transgenic crops is a big challenge for the success of disease resistant papaya crops. The aim of this review is to understand the genomic nature of PRSV, detection methods and the different advanced control methods. This review article will be helpful in developing the best management strategies for controlling PRSV.     

Identification of one novel causative mutation in exon 4 of WFS1 gene in two Italian siblings with classical DIDMOAD syndrome phenotype

The aim of the present paper is to describe a novel missense mutation (G107R) of WFS1 gene that was unexpectedly detected, in two siblings from Southern Italy, outside exon 8; a very unusual finding which has previously been reported only twice in Italian patients with Wolfram syndrome (WS). Although in Spanish pedigrees' WFS1 mutations are frequently located in exon 4, this finding is very infrequent in other pedigrees, particularly in Italian patients. Conclusions: a) our report of two siblings with one novel WSF1 mutation (G107R) expands the molecular spectrum of WS; b) this is the 3rd report of Italian patients harbouring one mutation outside exon 8 and the 2nd with one mutation in exon 4; c) on the basis of the present observations, and literature data we can infer that mutation locations outside exon 8 do not seem to be clearly associated with peculiar phenotype expressions of WFS1 gene.     

Genomic resources in fruit plants: an assessment of current status

The availability of many genomic resources such as genome sequences, functional genomics resources including microarrays and RNA-seq, sufficient numbers of molecular markers, express sequence tags (ESTs) and high-density genetic maps is causing a rapid acceleration of genetics and genomic research of many fruit plants. This is leading to an increase in our knowledge of the genes that are linked to many horticultural and agronomically important traits. Recently, some progress has also been made on the identification and functional analysis of miRNAs in some fruit plants. This is one of the most active research fields in plant sciences. The last decade has witnessed development of genomic resources in many fruit plants such as apple, banana, citrus, grapes, papaya, pears, strawberry etc.; however, many of them are still not being exploited. Furthermore, owing to lack of resources, infrastructure and research facilities in many lesser-developed countries, development of genomic resources in many underutilized or less-studied fruit crops, which grow in these countries, is limited. Thus, research emphasis should be given to those fruit crops for which genomic resources are relatively scarce. The development of genomic databases of these less-studied fruit crops will enable biotechnologists to identify target genes that underlie key horticultural and agronomical traits. This review presents an overview of the current status of the development of genomic resources in fruit plants with the main emphasis being on genome sequencing, EST resources, functional genomics resources including microarray and RNA-seq, identification of quantitative trait loci and construction of genetic maps as well as efforts made on the identification and functional analysis of miRNAs in fruit plants.