水稻小粒矮突变性状与T-DNA插入标签的共分离检测

 从T-DNA水稻标签系中发现1个标签系中出现了小粒矮(sgd)突变性状的分离。为克隆该突变基因和进行基因功能研究,对该突变体进行了遗传分析。该标签系存在多个插入事件,且T3代发现目标突变性状分离系中,另一个非目标插入事件已经纯合,故特异PCR检测不能确认小粒矮突变性状与T-NA插入共分离。但通过TAIL-CR方法,获得了插入点侧翼的水稻基因组序列,并证实该侧翼序列与小粒矮突变性状共分离。     

Empirical estimation of recreational exploitation of burbot,Lota lota,in the Wind River drainage of Wyoming using a multistate capture–recapture model

Burbot, Lota lota (Linnaeus), is a regionally popular sportfish in the Wind River drainage of Wyoming, USA, at the southern boundary of the range of the species. Recent declines in burbot abundances were hypothesised to be caused by overexploitation, entrainment in irrigation canals and habitat loss. This study addressed the overexploitation hypothesis using tagging data to generate reliable exploitation, abundance and density estimates from a multistate capture–recapture model that accounted for incomplete angler reporting and tag loss. Exploitation rate μ was variable among the study lakes and inversely correlated with density. Exploitation thresholds μ₄₀ associated with population densities remaining above 40% of carrying capacity were generated to characterise risk of overharvest using exploitation and density estimates from tagging data and a logistic surplus‐production model parameterised with data from other burbot populations. Bull Lake (μ = 0.06, 95% CI: 0.03–0.11; μ₄₀ = 0.18) and Torrey Lake (μ = 0.02, 95% CI: 0.00–0.11; μ₄₀ = 0.18) had a low risk of overfishing, Upper Dinwoody Lake had intermediate risk (μ = 0.08, 95% CI: 0.02–0.32; μ₄₀ = 0.18) and Lower Dinwoody Lake had high risk (μ = 0.32, 95% CI: 0.10–0.67; μ₄₀ = 0.08). These exploitation and density estimates can be used to guide sustainable management of the Wind River drainage recreational burbot fishery and inform management of other burbot fisheries elsewhere.     

Food quality affects production of Lumbricus terrestris (L.) under controlled environmental conditions

Birch leaves and horse manure were used to determine the effects of food quality on growth and reproduction of laboratory-reared Lumbricus terrestris. Animals grew to maturity within 6 months but attained a significantly (p < 0.001) larger adult size with manure (6.17 g) versus leaves (4.20 g). Cocoon production by recently-mated adults maintained in isolation, fed with birch leaves or horse manure, resulted in 4.53 and 3.84 cocoons ind.⁻¹ month⁻¹ respectively, with an initial hatchability of 86%, falling to zero after 18 months. Re-mating of these known individuals permitted long term monitoring of reproductive output (to 30 months). For the whole experimental period, overall hatchability of the 2010 cocoons produced was 44.4%. Median incubation time of those cocoons that hatched within accepted norms (less than 5 months at 15 °C) was 103 days and was not influenced by adult food type. A proportion (35.5%) of cocoons took in excess of 12 months to hatch. Adult mortality was minimal (25%) during the long term experiment but abnormal cocoon production was recorded after 2 years. Overall results demonstrate that food quality can have a significant influence on somatic and reproductive production of L. terrestris and these data may aid construction of production models for this earthworm in ecosystems with contrasting food quality.     

Alteration of wheat vernalization requirement by alien chromosome-mediated transposition of MITE

Under the changing climate, early flowering is advantageous to escape terminal heat and drought. Previously during evaluation of 14 chromosome introgression lines (ILs), we found three ILs that flowered a month earlier than their wheat background Chinese Spring (CS). This paper describes the cause of the early flowering in the ILs and provides insight into the evolution of spring wheat from the winter wheat. We used specific molecular markers for Vrn genes to determine its allelic composition. Phenotypic evaluations carried out under field conditions and in a growth chamber. Unlike the winter vrn-A1 allele of CS, the spring Vrn-A1 allele of the ILs had insertions of 222 and 131-bp miniature inverted-repeat transposable element (MITE) in the promoter region. Sequence analysis indicated that the 222-bp insertion is similar to an insertion in the spring genotype, Triple Dirk D. Our results ruled out any possibility of outcrossing or contamination. Without vernalization, Vrn-A1 is highly expressed in the ILs compared to CS. We attribute the early flowering of the ILs to the insertion of the MITE in the promoter of Vrn-A1. The alien chromosome might mediate this insertion.     

Genetic basis of phenotypic differences between transplanted stocks of brown trout

Abstract– We present results from an experiment testing for the existence of genetically based phenotypic differences among populations of brown trout ( Salmo trutta L.) born and raised under entirely natural environmental conditions. Genetically tagged individuals from two stocks (A and B) were introduced into a drainage system in Sweden previously void of brown trout, and the first generation (F₁) progeny were sampled from two lakes during nine consecutive years. Phenotypic differences among groups of progeny (A, B, and the AB hybrid) are expected to reflect genetically determined dissimilarities between the introduced stocks. Phenotypic differences among progeny groups were observed for age at maturity and for migratory and reproductive behavior, and these characters are apparently determined by genetic factors to an extent that permit their detection even in the presence of confounding and naturally occurring sources of variation such as lake, age, cohort and year of sampling. There was also significant variation among offspring groups with respect to body size (length), but only a small proportion of the total variation in size could be attributed to stock differences. These genetically based stock characteristics may represent local adaptations, and the fishery management implications of these findings are discussed.     

The role of siderophores in potato tuber yield increase by Pseudomonas putida in a short rotation of potato

The effect of treatment of potato seed tubers withPseudomonas putida isolate WCS358 on tuber yield was studied in different crop rotations at the Experimental Farm De Schreef, near Lelystad. With untreated, tuber yield in a 1:3 (short) rotation compared to yield in a 1:6 (long) rotation of potato was decreased by 11% at 86 days (seed tuber harvest) and by 14% at 130 days (ware potato harvest) after seeding. Seed tuber treatment with the wild-type isolate WCS358 increased tuber yield with 13% in a short rotation of potato 86 days after seeding, whereas a siderophore-negative Tn5 transposon mutant of this isolate had no effect on tuber yield. Seed tuber treatment with the wild-type isolate or the siderophore-negative mutant in a long rotation of potato had no effect on tuber yield. At 130 days after seeding no effect of any of the seed tuber treatments was found in both short and long rotations of potato.Root colonization by siderophore-producing Tn5 transposon mutants of WCS358 was decreased at the end of the growing season. No difference in root colonization between siderophore-producing and siderophore-negative Tn5 transposon mutants was found at 130 days after seeding.Siderophore production seems to be a prerequisite in potato tuber yield increase by WCS358 under field conditions. This is the first time that the involvement of siderophores in growth stimulation has been demonstrated in the field.Samenvatting De invloed van een behandeling van aardappelpootgoed metPseudomonas putida isolaat WCS358 op de knolopbrengst werd onderzocht in verschillende gewasrotaties on een proefveld van proefboerderij De Schreef, Flevopolder. In de controlebehandelingen werd in een nauwe aardappelrotatie (1:3) een reductie van 11% in opbrengst van pootaardappelen (86 dagen na het poten) geconstateerd ten opzichte van een ruime aardappelrotatie (1:6); 130 dagen na het poten werd een vermindering met 14% gevonden in de opbrengst van consumptieaardappelen.Pootgoedbehandeling met het siderofoorproducerende isolaat WCS358 verhoogde de opbrengst van pootaardappelen in de 1:3-rotatie met 13%. Een Tn5-transposonmutant van dit isolaat die het vermogen sideroforen te produceren had verloren, had geen effect op de opbrengst. In de 1:6-rotatie had behandeling van pootgoed met WCS358 geen effect op de opbrengst van pootaardappelen.Zowel in de nauwe (1:3) als in de ruimte (1:6) rotatie werd (130 dagen na het poten), geen effect van behandeling van pootgoed met WCS358 op de opbrengst van consumptieaardappelen gevonden.Wortelkolonisatie door siderofoorproducerende Tn5-transposonmutanten van WCS358 nam aan het eind van het seizoen af. Er werd, 130 dagen na het poten, geen verschil in wortelkolonisatie geconstateerd tussen siderofoorproducerende en siderofoornegatieve Tn5-transposonmutanten.Siderofoorproduktie blijkt een voorwaarde te zijn voor verhoging van de knolopbrengst door WCS358 onder veldomstandigheden. De verhoging van de knolopbrengst treedt alleen op in de nauwe aardappelrotatie. Dit is de eerste keer dat de betrokkenheid van sideroforen bij groeistimulatie onder veldomstandigheden is aangetoond.     

转座子标签法克隆水稻基因前景

转座子标签法克隆基因是近年来发展起来的一种非常有效的分子生物技术。介绍了转座子标签技术克隆基因的基本原理及研究进展,并对转座子标签技术在水稻基因克隆上的应用进行了讨论分析。     

一个链霉菌中微型转座子系统的构建

针对链霉菌中现有转座子系统的一些缺陷,构建了一个链霉菌中的微型转座子质粒pHL265,它携带的转座酶基因tnpA在转座子mini-Tn4560A外部,降低了发生二次转座的可能性,并带有大肠杆菌与链霉菌进行属间接合转移的起始位点oriT,可通过接合转移的方式将其从大肠杆菌导入到链霉菌中.利用该转座子系统转座筛选得到天蓝色链霉菌M145的约1 000株转座突变株.选取其中的8株,经Southern杂交验证可知,该转座子的转座基本具有随机性,并在宿主DNA中稳定存在.此系统为链霉菌功能基因组的研究提供了新的技术手段.     

利用细菌转座子构建适于家蚕的Bac-to-Bac快速基因表达系统

为了较好地解决家蚕杆状病毒基因表达系统(BmNPV expression system)在重组病毒构建和纯化过程中存在的重组率低、空斑分析技术繁琐、花费时间长等缺点,借鉴国外AcNPV Bac-to-Bac快速基因表达系统工作原理,对家蚕BmNPV基因组进行了改造。通过同源重组的方法,将含有单拷贝数细菌F复制子、插入有细菌转座子整合靶位点、编码LacZα肽的部分DNA片段和抗性选择标记基因的基因片段重组入家蚕BmNPV基因组,替换多角体蛋白基因,获得了家蚕BmNPV病毒穿梭载体BmBacm id;并利用供体质粒上的表达盒和细菌转座子以及细菌转座子的基因定位转移作用,在细菌体内实现外源目的基因向家蚕BmNPV基因组上的转移整合,快速完成重组BmN-PV病毒的构建。     

Tn5 transposon mutagenesis in Acidovorax citrulli for identification of genes required for pathogenicity on cucumber

An Acidovorax citrulli–cucumber pathosystem was established through which A. citrulli mutants with altered pathogenicity, generated by transposon mutagenesis, were identified on cucumber cotyledons. The A. citrulli group I strain FC440 was shown to grow faster in cucumber leaf tissues than a group II strain and was used for Tn5 transposon mutagenesis. A total of 2100 Tn5 insertional mutants were generated, and analysis of the mutant library showed that the transposon insertions were single, independent and stable. A conserved non‐flagellar type III secretion system (NF‐T3SS) ATPase gene hrcN was identified and confirmed to be essential for pathogenicity and functionality of NF‐T3SS in A. citrulli. Comparative sequence analysis of the HrcN protein and its homologues in other representative bacterial plant pathogens revealed that the NF‐T3SS of A. citrulli is close to that of Ralstonia solanacearum and Xanthomonas campestris, but distant from that of Pseudomonas syringae and Erwinia amylovora. The generated Tn5 insertional mutant collection is valuable for identification of genes required for A. citrulli pathogenesis, and the established A. citrulli–cucumber pathosystem will facilitate an improved understanding of A. citrulli biology and pathology.