农杆菌介导陆地棉棕彩选1号茎尖遗传转化

为建立高效的棕色棉茎尖遗传转化体系,以陆地棉棕彩选1号为材料,采用农杆菌介导茎尖的遗传转化方法,以培养7 d的棕彩选1号幼苗茎尖为受体,设计正交试验对共培养时间、共培养温度、菌液OD600、侵染时间等条件进行优化;此外,还研究了受体茎尖的大小及真空渗透等因素对转化效率的影响,初步建立了棕色棉茎尖农杆菌介导的遗传转化技术体系。结果表明,带有0.5 cm下胚轴的茎尖适合转化,适宜的农杆菌介导遗传转化条件为农杆菌菌液浓度OD600=1.0,侵染时间20 min,共培养时间3 d,共培养温度24℃。此外,真空渗透处理有助于抗性苗的发生。再生植株经在含100 mg·L~(-1)卡那霉素的培养基上筛选,获得102株抗性植株,再进行PCR鉴定和Southern杂交检测,共获得24株转基因植株,转化率为0.4%,且为单拷贝基因插入。本试验建立的优化条件为棕色棉的遗传转化奠定了理论基础。     

根癌农杆菌介导转褪黑素基因草莓的获得

为建立根癌农杆菌介导的草莓褪黑素基因高效遗传转化体系,以草莓品种“早红”叶片和叶柄为外植体,优化了影响根癌农杆菌遗传转化的因素,包括农杆菌菌液浓度、侵染时间、共培养时间和预培养时间等,建立了适宜草莓“早红”的农杆菌遗传转化体系。结果表明,“早红”草莓适宜的转化条件是：外植体的最适预培养时间为2～3 d,农杆菌侵染的最佳菌液浓度为OD600 0.3～0.5,最适侵染时间为15 min,最适共培养时间为3 d。在此基础上,通过根癌农杆菌介导法将褪黑素合成关键酶N -乙酰转移酶（AANAT）和羟吲哚-氧甲基转移酶（HIOMT）基因AANAT和HIOMT转入草莓基因组,并对获得的抗性植株进行PCR和PCR-Southern blot检测分析,结果表明外源基因已经整合进草莓基因组中。RT-PCR分析证明外源基因已经在转录水平表达。     

农杆菌介导高羊茅遗传转化体系的建立

为建立农杆菌(Agrobacterium tumefaciens)介导高羊茅(Festuca arundinaceaSchreb.)遗传转化体系,以草坪型高羊茅品种凌志的胚性愈伤组织为受体材料,通过gus基因瞬时表达检测,研究了多种因素对农杆菌介导转化的影响。结果表明,农杆菌介导高羊茅胚性愈伤组织转化的适宜条件为:愈伤组织在含BAP和NAA各0.5mg/L的培养基上预培养3d;菌液浓度OD600值0.7,感染时间15min;农杆菌预培养基和悬浮培养基以及共培养基中添加100μmol/L的乙酰丁香酮;共培养温度23℃,共培养时间3d,共培养基pH值5.2。不同浓度潮霉素筛选效果试验表明,采用低浓度(50mg/L)潮霉素连续筛选,再生获得的转基因植株数最多,因此是最为可取的筛选方式。     

根癌农杆菌介导的蓝猪耳遗传转化

报道了根癌农杆菌（Agrobacterium tumifaciens）介导的蓝猪耳（Torenia foumieri）遗传转化的方法。筛选蓝猪耳转化芽的最适卡那霉素（Kan）浓度为400mg／L；以稀释10倍的菌液浸染叶盘10～20min，固体共培养基（含20μmol／L乙酰丁香酮）上23℃共培养7～8d可获得转化芽诱导率27．2％；16h光照，8h黑暗是较理想的共培养光周期；茎段作为外植体，其转化芽诱导率要高于叶盘的转化芽诱导率，而且其转化芽比叶盘的转化芽健壮；1／2MS＋100mg／L Kan＋0．5mg／L IAA是比较理想的生根培养基。实验结果表明，根癌农杆菌介导的蓝猪耳转化获得了抗性再生植株，转化率为24．1％。     

高粱茎尖再生体系及其遗传转化影响因子的研究

本文对高粱茎尖再生体系及其转化影响因子进行了研究。不同基因型的茎尖再生频率有很大差异,说明基因型是影响茎尖再生的一个重要因素。在9种不同的激素组合中,以0.25MG/L KT、0.5MG/LBAP和0.25MG/L IAA对茎尖的再生效果最好。在MS培养基的基础上,采用B5培养基维生素并添加200MG/L L-ASN,10MG/L抗坏血酸和100MG/L PVP可提高茎尖再生频率。对农杆菌介导高粱茎尖转化的因子进行了优化,获得较高转化频率的条件为:茎尖预培养时间3D,在OD600值为0.5农杆菌菌液中感染10MIN,共培养3D。     

农杆菌浸种法介导中华结缕草遗传转化体系的建立

以中华结缕草萌动种子为转化受体,利用NPT-Ⅱ筛选基因对影响农杆菌介导的遗传转化外部因素进行探索,通过优化转化条件,建立了农杆菌浸种途径介导的中华结缕草遗传转化体系。结果表明：利用催芽6-7d的种子,加入OD600值为0.9的农杆菌浸染菌液（在浸染菌液中添加100μmol.L-1乙酰丁香酮）,附加5min+30s真空处理,振荡浸染48h,共培养3d后,用75mg.L-1卡那霉素筛选能够获得较好的转化效果。通过对卡那霉素抗性苗进行PCR检测和GUS组织化学染色,初步验证NPT-Ⅱ和GUS基因已经整合进植物基因组,阳性转化率为55.21%。     

抗生素对高粱茎尖再生的影响及再生体系的建立

研究羧苄青霉素、头孢霉素、潮霉素和卡那霉素4种抗生素对高粱茎尖再生的影响及再生体系的建立结果表明,MS培养基中添加甘氨酸、乙二胺四乙酸的铁络合物(Fe-EDTA)和抗坏血酸等有机物有利于高粱茎尖的分化与生长。农杆菌介导的高粱茎尖遗传转化过程中羧苄青霉素为最理想抗生素,最佳浓度为250mg／L。并初步确立了高粱茎尖农杆菌介导的遗传转化过程中抑菌和转化体的筛选策略。     

抗生素对高粱茎尖再生的影响及再生体系的建立

研究羧苄青霉素、头孢霉素、潮霉素和卡那霉素4种抗生素对高粱茎尖再生的影响及再生体系的建立结果表明,MS培养基中添加甘氨酸、乙二胺四乙酸的铁络合物(Fe EDTA)和抗坏血酸等有机物有利于高粱茎尖的分化与生长。农杆菌介导的高粱茎尖遗传转化过程中羧苄青霉素为最理想抗生素,最佳浓度为2 5 0mg/ L。并初步确立了高粱茎尖农杆菌介导的遗传转化过程中抑菌和转化体的筛选策略     

农杆菌介导春小麦遗传转化体系的优化研究

采用携带GUS基因双元表达载体p1301-Pubi-Ecppc的根癌农杆菌菌株C58c1、LBA4404和EHA105对4个春小麦品种的幼胚愈伤组织进行了遗传转化。结果表明,3M1.5为最佳诱导培养基;扬麦158和扬麦10是用于小麦遗传转化的良好基因型。对农杆菌转化条件进行进一步优化,得出用农杆菌C58c1侵染预培养15d的扬麦158幼胚愈伤组织,当侵染液浓度为OD6000.8、侵染40min、共培养3d时,可以提高农杆菌介导小麦遗传转化的筛选频率和PCR阳性率。对所得到的具有hyg抗性的愈伤和抗性植株进行GUS基因化学组织检测和PCR分子鉴定,初步证明Ecppc基因已整合到小麦再生植株基因组中。本研究初步建立了农杆菌介导小麦的遗传转化体系。     

棉花农杆菌基因转化体系的优化和抗黄萎病转基因植株的获得

为了优化根癌农杆菌介导的棉花下胚轴切段的基因转化体系,以苜蓿抗菌肽基因（alflafa antifungul peptide gene, alfAFP）为目标,利用GUS报告基因和潮霉素抗性基因的检测方法,分析和探讨了根癌农杆菌菌种、菌液浓度、浸染下胚轴的时间、共培养中乙酰丁香酮（AS）的浓度、共培养温度和时间等因子对基因转化的影响。优化分析表明,用农杆菌菌株LBA4404当OD600值为0.5-0.7的菌液浸染5-7日龄的棉花无菌苗下胚轴切段10-15分钟,在含200µmol/L AS的共培养基中21℃暗培养60h可更有效地提高转化率。经上述条件处理的下胚轴在含50mg/L潮霉素的愈伤诱导培养基上培养3周可产生转化率最高的愈伤。愈伤组织经分化培养获得15株再生棉花,经过PCR和southern-PCR分析,发现其中12株含有外源抗病基因alfAFP,可作为棉花抗病育种的种质。     

Potential of wild species for genetic enhancement of some semi-arid food crops

Discovery and incorporation of genes from wild species provide means to sustain crop improvement, particularly when levels of resistance in the cultigens are low and virulent strains of pests and pathogens overcome the host plant resistance. The extent of utilization and the potential of the wild genepool for genetic enhancement were reviewed in five important food crops viz. sorghum, pearl millet, chickpea, pigeonpea and groundnut grown in the semi-arid tropics. Introgression from compatible wild germplasm in the primary gene pool resulted in transfer of new cytoplasmic male sterility systems in pearl millet and pigeonpea, development of high protein, cleistogamous flower and dwarf pigeonpea lines and foliar disease resistant groundnut cultivars. Utilization of wild species in secondary and tertiary gene pools has been generally limited due to sterility, restricted recombination or cross incompatibility. Nevertheless, these species are extremely important as they contain high levels of resistance to several important biotic and abiotic stresses. Several of them, like those belonging to the Parasorghum section and the rhizomatous Arachis species are sources of multiple resistances and hold great promise to sustain crop productivity.     

“绿色革命”以来大气CO2浓度升高对C3作物营养品质的影响

自20世纪60年代"绿色革命"以来,育种技术和农耕技术的发展促进了农作物产量的大幅提升,然而作物的营养品质出现下降趋势。在相似的遗传背景下,大气CO_2浓度升高会使单位体积农作物产品的营养元素含量下降,因此"绿色革命"至今,农作物产品的营养元素下降可能受大气CO_2浓度升高影响。通过植物生长箱模拟"绿色革命"初期和目前的大气CO_2浓度水平(310μmol/mol和400μmol/mol),针对主要C_3作物水稻、小麦和大豆,研究"绿色革命"以来大气CO_2浓度升高对其籽粒的C、N、Fe、Zn元素含量的影响,结果表明:CO_2浓度升高对3种作物籽粒的C元素含量几乎没有影响,变化幅度在±1.5%之间;籽粒的N、Fe、Zn元素含量普遍呈现下降趋势,但均未达到显著水平。     

Faculty positions:Center for Agricultural Resources Research,Chinese Academy of Sciences

正The Center for Agricultural Resources Research(CARR),the Institute of Genetics and Developmental Biology(IGDB),Chinese Academy of Sciences,invites applicants for several research group leader positions.CARR is one of the research organizations in Chinese Academy of Sciences(CAS).We seek nominations and applications     

Collection and preliminary evaluation of native turfgrass accessions in Italy

Despite a raising interest on turfs in Italy, all theavailable varieties of this kind in the Country are of foreign origin, and areoften poorly adapted to the prevailing climatic conditions. This prompted tobegin a collection activity of indigenous turfgrass species, with the ultimategoal of identifying promising materials for future breeding based on localgenetic resources. The collection was carried out in three areas of Italy, viz.the northern Po Plain, the coastal region of Liguria, and the island of Sardiniathat are characterised, respectively, by subcontinental, warm temperate, andtypical Mediterranean climate. Altogether, 141 sites were visited, yielding 226accessions belonging to eight species of potential interest for turfs:Poa pratensis, Poa trivialis,Festuca rubra, Festuca arundinacea,Lolium perenne, Agrostis stolonifera,Agrostis tenuis, and Cynodon dactylon,this last being a warm-season grass. Poa pratensis andCynodon dactylon were mostly collected in northern Italyand Sardinia, respectively, whereas Festuca arundinacea andLolium perenne were rather ubiquitous. The collection sitesranged from 0 to 1040 m asl, but sites over 750 m wereonly visited in the inner part of Sardinia. All the accessions, collected aswhole plants, were transplanted at Lodi, northern Italy, where they are beingevaluated. Their preliminary evaluation for traits of importance for turf use,such as sward colour and overall quality, highlighted the great variation andthe occurrence of interesting accessions in all species. Other characters wererecorded, bearing specific importance in individual species, and in all casespromising accessions were identified. The germplasm of Festucarubra, Festuca arundinacea, and Loliumperenne proved highly infected by endophytic (symbiotic) fungi of thegenus Neotyphodium.     

Leaf position and genotype differences for mineral element concentrations in sorghum grown on tropical acid soil 1

Mineral element deficiencies and toxicities are common problems associated with sorghum [Sorghum bicolor (L.) Moench] production on acid soils. To better understand some of the mineral element problems and the analysis of plant tissue of sorghum plants grown on acid soils, four sorghum genotypes were grown on an acid Oxisol at Carimagua, Colombia limed with dolomite at 2 and 6 Mg ha^‐1.

Samples for mineral element analyses were obtained from leaves at different positions on the four genotypes. Concentrations of P and Mg were highest in the flag leaf (Leaf No. 1) and decreased as the position on the plant declined from the top of the plant for plants grown at 2 Mg lime ha^‐1. Similar decreases in P, Mg, K, and Zn concentrations occurred in plants grown with 6 Mg lime ha^‐1. Concentrations of Ca, S, Si, Mn, Fe, Cu, and Al increased as leaf position declined from the flag leaf for plants grown at 2 and 6 Mg lime ha^‐1. The higher lime supply enhanced Ca and reduced Mn and Fe concentrations in leaves. Differences in mineral element concentrations for the four genotypes used were fairly extensive. The elements to show the greatest range among genotypes were Al and Si and the elements to show the least range among genotypes were P, K, and S. Care should be used in collecting leaf samples for plant analysis and genotypic differences for accumulation of mineral elements should be considered in interpretation of results.     

Short-term variation in the concentration of selected ions within individual tropical rainstorms

Rainwater was collected at the campus of the University of Brunei Darussalam in Bandar Seri Begawan, Brunei Darussalam, using a funnel-in-bottle sampler. Polypropylene bottles were changed at intervals during rainstorm events. The pH and conductivity were determined immediately after collection on aliquots of the sample. Samples were refrigerated at 5°C for subsequent chemical analysis. Analyses for Na, Mg, Ca, Zn and Fe were carried out by means of inductively coupled plasma atomic emission spectroscopy (ICP-AES); Cu and Mn were analysed by graphite furnace atomic absorption spectroscopy (GFAAS); K was analysed using flame atomic emission spectroscopy (FAES); and Cl^–, NO₃ ^– and SO₄ ^2– were analysed by ion chromatography (IC). Concentration versus time profiles are reported for three rainstorm events. All ions exhibited a decrease in concentration during the rainstorm. The first sample contained the highest concentration of ions, consistent with a first-flush effect. The contribution of the initial stages of the shower to the total quantity of ion deposited during the entire rainstorm is quite overwhelming; in many cases 20 to 30% of the mass was deposited in less than 5% of rainstorm duration. On the other hand, the pH and conductivity variation during rainstorms did not exhibit a consistent pattern.     

Leaf litter selection by detritivore and geophagous earthworms

Summary Litterbag experiments with 10 different kinds of leaf litter showed that detritivore (Lumbricus species) and geophagous (Aporrectodea species) earthworms prefer certain litter types over others, since different numbers of worms were found below the litter after 50–52 days of exposure in a pasture. The detritivores preferred Fraxinus, Tilia, and predecomposed Ulmus and Fagus litter to Fagus litter and paper, while geophages preferred Tilia litter to Alnus and Ulmus litter, so that the two groups of earthworms showed different preferences. The detritivores seemed to be more selective than the geophages. The palatability of the litter was examined in relation to the C: N ratio, the lignin concentration and the initial and final polyphenol concentration. The numbers of detritivores were significantly correlated with the C: N ratio and the final polyphenol concentration, so that selection of litter seems to be related to palatability. The numbers of geophages were not significantly correlated with any of the parameters for palatability. The disappearance of litter after 50–52 days appeared to be due to detritivore activity, since the numbers found below the litter were positively and significantly correlated with the litter disappearance. There was no significant correlation with geophage activity. This indicates that detritivores use litter as food, and therefore influence the composition of the litter layer.     

The 5S rRNA gene diversity in the genus <Emphasis Type="Italic">Secale</Emphasis> and determination of its closest haplomes

We analyzed 127 rDNA sequences (5S DNA units) obtained from 23 seed accession samples from more or less 10 taxa in wild and cultivated rye, genus Secale L. The sequences fell into two known groups, here assigned to two unit classes, viz. long R1 and short R1 (designations to reflect on R haplome of rye). The different taxa could not be fully differentiated based on the 5S DNA units. We searched for 5S DNA sequences from known unit classes most closely similar to the long R1 and the short R1. One set with the long R1 unit class contained sequences of the long P1 unit class from Agropyron (P haplome) and from Kengyilia (StYP haplome), long J1 from Thinopyrum (J haplome), whereas the set with the short R1 included the long S1 from Pseudoroegneria (St haplome) and Kengyilia (StYP haplome), the short J1 from Thinopyrum (J haplome) and the short V1 from Dasypyrum (V haplome). Each of the two sets was analyzed separately by maximum likelihood (ML) phylogenetic analysis from which we were able to infer that the 5S DNA units of Secale differentiated in a non-clock fashion and followed the HKY substitution model in the gene tree with the long R1 unit class and the HKY + G in the gene tree with the short R1 unit class. A complementary Bayesian analysis yielded identical tree topologies to the ML ones for each of the two sequence sets. In the tree with the long R1 units the long P1 and long J1 unit classes were closest to the long R1 unit class, whereas in the tree with the short R1 units the long S1 and short J1 unit classes were closest to the short R1 unit class, indicating possibly a close relationship between the St, J and R haplomes.     

Nematode succession and microfauna-microorganism interactions during root residue decomposition

The quality of plant material affects the vigor of the decomposition process and composition of the decomposer biota. Root residues from hairy vetch (Vicia villosa Roth), rye (Secale cereale L.) and vetch+rye, packed in litterbags were placed in pots of soil at 15 C and the content of the bags was analyzed after 2, 4, 8 and 12 weeks. Bacterial biomass did not differ between residues with contrasting composition. Among bacterivores groups of nematodes that require high bacterial production dominated in fast decomposing resources whereas flagellates with smaller requirements prevail in slower decomposing resources. Biomass of bacterial feeding nematodes correlated positively with early phase (0-2 wk) decomposition that increased in the order: rye< vetch+rye<vetch. Bacterial biomass therefore seems to be under top-down (predation) control during early decomposition. In contrast, the fungal biomass differed between resources with highest values for rye. Moreover, this increase in fungal biomass occurred later during succession and was correlated with decomposition activity for rye in that period. Fungal biomass therefore seems to be under bottom-up (resource) control. The composition of the nematode assemblages (composed of 25 taxa) showed a clear relationship to initial plant resource quality as well as decomposition phase. Early successional microbivorous nematodes vary according to resource quality with demanding bacterivores+predators (Neodiplogasteridae) dominating in vetch and less demanding bacterivores (Rhabditidae) and fungivores (Aphelenchus) being equally common in vetch and rye. Later in the succession (2-4 wk) bacterivorous Cephalobidae and fungivorous Aphelenchoides prevailed similarly on the different root materials whereas bacterivorous protozoa and the amoebal fraction thereof dominated in rye. At week 12 no species dominated the nematode assemblages that were similar between the resources. The differences between nematode assemblages among plant resources at 2 week were similar to the results of a field study sampled after 6 weeks with the same soil and plant resources. This lends support to the relevance of the successional patterns observed in this incubation study.     

Cycling of extracellular DNA in the soil environment

Upon entering the soil environment, extracellular DNA is subjected to dynamic biological, physical, and chemical factors that determine its fate. This review concerns the fate of both recombinant and non-recombinant sources of DNA. A schematic of DNA cycling coupled with genetic transformation is presented to understand its behavior in soil. Extracellular DNA may persist through cation bridging onto soil minerals and humic substances, be enzymatically degraded and restricted by DNases of microbial origin, and/or enter the microbial DNA cycle through natural transformation of competent bacteria. Lateral gene transfer may disseminate DNA through the microbial community. An understanding of DNA cycling is fundamental to elucidating the fate of extracellular DNA in the soil environment.