Phyto-availability and speciation change of heavy metals in soils amended with lignin as micro-fertilizer

Lignin is a primary byproduct from the black liquor treatment in paper making industries, its application as micro-fertilizer in agricultural land might provide a promising alternative to sewage discharge. However, application of such a micro-fertilizer might affect the soil properties and result in soil pollution. In this study, the effects of lignin application on phyto-availability and speciation change of heavy metals in soils were investigated. Greenhouse experiments showed that lignin application improved the growth of winter wheat (Triticum aestivum L.) in all three soils investigated. The increase of the biomass for wheat shoot was 59.7%, 39.8% and 12.3% for Beijing soil, Jiangxi soil and Dongbei soil, respectively. In contrast, lignin amendment decreased the concentrations of heavy metals in wheat shoots from 2.2% to 61.0%. Sequential extraction procedure of a three-step BCR was used to investigate the fraction distribution. The extractable fractions were specified as fraction B1: water soluble,     

Recycling Isolation of Plant DNA,A Novel Method

DNA is one of the most basic and essential genetic materials in the field of molecular biology.To date,isolation of sufficient and good-quality DNA is still a challenge for many plant species,though various DNA extraction methods have been published.In the present paper,a recycling DNA extraction method was proposed.The key step of this method was that a single plant tissue sample was recycled for DNA extraction for up to four times,and correspondingly four DNA precipitations(termed as the 1st,2nd,3rd and 4th DNA sample, respectively) were conducted.This recycling step was integrated into the conventional CTAB DNA extraction method to establish a recycling CTAB method.This modified CTAB method was tested in eight plant species,wheat,sorghum,barley,corn,rice,Brachypodium distachyon,Miscanthus sinensis and tung tree.The results showed that high-yield and good-quality DNA samples could be obtained by using this new method in all the eight plant species.The DNA samples were good templates for PCR amplification of both ISSR and SSR markers.The recycling method can be used in multiple plant species and can be integrated with multiple conventional DNA isolation methods,and thus is an effective and universal DNA isolation method.     

A Neuro-Muscular Elasto-Dynamic Model of the Human Arm Part 1: Model Development

In this paper we develop an elasto-dynamic model of the human arm for use in neuro-muscular control and dynamic interactionstudies.The motivation for this work is to present a case for developing and using non-quasistatic models of humanmusculo-skeletal biomechanics.The model is based on hybrid parameter multiple body system(HPMBS)variational projectionprinciples.In this paper,we present an overview of the HPMBS variational principle applied to the full elasto-dynamic model ofthe arm.The generality of the model allows one to incorporate muscle effects as either loads transmitted through the tendon atpoints of origin and insertion or as an effective torque at a joint.Though the technique is suitable for detailed bone and jointmodeling,we present in this initial effort only simple geometry with the bones discretized as Rayleigh beams with elongation,while allowing for large deflections.Simulations demonstrate the viability of the mcthod for use in the companion paper and infuture studies.     

小麦雄性不育遗传及基因定位研究进展

雄性不育的研究对于杂种优势的利用具有重要意义。本文综述了小麦雄性不育遗传及基因定位研究进展,介绍了小麦雄性不育的基因工程,对小麦雄性不育的应用进行了讨论。 Abstract:The study of plant male sterility plays an important role on utilization of heterosis.This paper reviews the current status of the studies of the heredity and mapping of the male sterile genes in wheat and the gene engineering of wheat male sterility.The application of male sterility in wheat breeding is discussed.     

Rapid detection of Shigella and Salmonella in rhesus monkeys by loop-mediated isothermal amplification assay

Objective To establish a loop-mediated isothermal amplification( LAMP) method for detecting diarrhea pathogens( Shigella and Salmonella) in rhesus monkeys and evaluate the application of the LAMP method for detecting bacterial diseases in nonhuman primate laboratory animals. Materials and Methods A total of 205 fecal samples of rhesus monkeys were detected in this LAMP assay. The specificity and sensitivity of LAMP for Shigella and Salmonella were analyzed,and real-time polymerase chain reaction( REAL-TIME PCR) assay was employed as control. Results The LAMP method established here needed only 45 min to complete the reaction at 63℃. Its detection limit was 10 pg / μL and with a high specificity. The positive rate of Shigella and Salmonella was 1. 5% and 6. 3%,respectively. Conclusions Here we have established a fast and simple Shigella and Salmonella LAMP detection method that has strong specificity and high sensitivity and is suitable for rapid detection of bacterial disease in macaques. The development of this rapid detection kit is underway,and it will be helpful to the diarrhea detection.     

Reproduction strategy of Trias verrucosa (Orchidaceae) from China

Trias, an orchid genus, was reported in this paper from China as a newly recorded genus for the first time. The genus and its new species, T. verrucosa Z. J. Liu, L. J. Chen et S. P. Lei, were described and discussed. Trias consists of 19 species ranging from Assam of India to Indonesia and Australia in the southeast via Myanmar to Deccan of India in the southwest. Its occurrence in West Yunnan of China is a further indication of phytogeographical relations between this region and the tropical Asia. This new species grows on shady rocks in forests. By observing of its biological characteristics such as phenology and blooming biology, lots of cloned ramets of generation overlap were found, but there were no fruited plants, and that clonal reproduction could repeat had nothing to do with whether the ramet bloomed or not. The flowering season of this species is from early April to early May. The opening of flowers on plants or inflorescence was irregular and the florescence of single flowers was rather short, only lasting 4–5 d, and no fruited flowers were found. Based on the detection of mating systems, no flowers of artificial self-pollination and artificial cross-pollination have fruited. Since the flowering period is just before the rainy season in this region, the ecological conditions of dryness, strong wind, low temperature and weak light during that period of time are evidently unfavorable to plant blooming, pollinating and fruiting. Apparently, the short duration and sterility of each flower would avoid the invalid energy waste in unfavorable circumstances and save the limited energy for more valid asexual reproduction so that the opportunity of multiplication in an uncertain environment would be increased to ensure the progenitive success. The P/O value of T. verrucosa is 187.4 ± 22.4, which is obviously related with the highly valid usage of its conglutinated and hard tuberous pollinarium during pollination, indicating that this species could hold the characteristic of sexual reproduction. This is an adaptation of this type of plants to the rather atrocious ecological circumstance there by its reproduction strategy of strengthening asexual reproduction and weakening sexual reproduction, and so it enables the plants to survive in this region, long-distanced from the places where its relatives dwell.     

Reproduction strategy of Trias verrucosa (Orchidaceae) from China

Trias, an orchid genus, was reported in this paper from China as a newly recorded genus for the first time. The genus and its new species, T. verrucosa Z. J. Liu, L. J. Chen et S. P. Lei, were described and discussed. Trias consists of 19 species ranging from Assam of India to Indonesia and Australia in the southeast via Myanmar to Deccan of India in the southwest. Its occurrence in West Yunnan of China is a further indication of phytogeographical relations between this region and the tropical Asia. This new species grows on shady rocks in forests. By observing of its biological characteristics such as phenology and blooming biology, lots of cloned ramets of generation overlap were found, but there were no fruited plants, and that clonal reproduction could repeat had nothing to do with whether the ramet bloomed or not. The flowering season of this species is from early April to early May. The opening of flowers on plants or inflorescence was irregular and the florescence of single flowers was rather short, only lasting 4–5 d, and no fruited flowers were found. Based on the detection of mating systems, no flowers of artificial self-pollination and artificial cross-pollination have fruited. Since the flowering period is just before the rainy season in this region, the ecological conditions of dryness, strong wind, low temperature and weak light during that period of time are evidently unfavorable to plant blooming, pollinating and fruiting. Apparently, the short duration and sterility of each flower would avoid the invalid energy waste in unfavorable circumstances and save the limited energy for more valid asexual reproduction so that the opportunity of multiplication in an uncertain environment would be increased to ensure the progenitive success. The P/O value of T. verrucosa is 187.4 ± 22.4, which is obviously related with the highly valid usage of its conglutinated and hard tuberous pollinarium during pollination, indicating that this species could hold the characteristic of sexual reproduction. This is an adaptation of this type of plants to the rather atrocious ecological circumstance there by its reproduction strategy of strengthening asexual reproduction and weakening sexual reproduction, and so it enables the plants to survive in this region, long-distanced from the places where its relatives dwell.     

Mapping of a BYDV resistance gene from Thinopyrum intermedium in wheat background by molecular markers

The wheat line H960642 is a homozygous wheat-Thinopyrum intermedium translocation line with resistance to BYDV by genomie in situ hybridization (GISH) and RFLP analysis. The genomie DNA of Th. intermedium was used as a probe, and eonunon wheat genomie DNA as a blocking in GISH experiment. The results showed that the chromosome segments of Th. intermedium were transferred to the distal end of a pair of wheat chromosomes. RFLP analysis indicated that the transloeation line H960642 is a T7DS·7DL-7XL translocation by using 8 probes mapped on the homoeologous group 7 in wheat. The tranalocation breakpoint is located between Xpsr680 and Xpsr965 about 90—99 cM from the centromere. The RFLP markers psr680 and psr687 were closoly linked with the BYDV resistance gene. The gene is located on the distal end of 7XL around Xpsr680 and Xpsr687.     

Structural and Expressional Variation Analyses of Mitochondrial Genomes Reveal Candidate Transcripts for the SV Cytoplasmic Male Sterility in Wheat (Triticum aestivum L.)

Common wheat (Triticum aestivum L.) is one of the most important crops,and intra-specific wheat hybrids have obvious heterosis in yield and protein quality.Therefore,utilization of hybrid wheat varieties offers an effective way to increase yield and nutrition.Cytoplasmic male sterility (CMS) systems are a useful genetic tool for hybrid crop breeding,and are ideal models for studying the genetic interaction and cooperative function of mitochondrial and nuclear genomes in plants (Schnable and Wise,1998;Hanson and Bentolila,2004).The breeding of hybrid wheat using male sterility caused by the cytoplasm of T.timopheevii has been studied since the early 1960's.But it is unsuccessful because there are some deficiencies in the practical application of this cytoplasm,including limited restoration resources,thin seeds,pre-harvest sprouting and lower germination rate (Wilson and Ross,1962).The Sv type of cytoplasmic male sterility (CMS-Sv) in wheat is general accessions for four types of CMS lines that were derived from four Aegilops species:Ae.kotschyi,Ae.variabilis,Ae.ventricosa,and Ae.bicornis.Based on the observation of alloplasmic lines produced in all possible combinations between 12 wheat nuclear genotypes and 47 cytoplasms of two related genera,Triticum (wheat) and Aegilops,Ogihara and Tsunewaki (1988) concluded that the D2-cytoplasm of Ae.crassa and its relatives,N-cytoplasm of Ae.uniaristata,and SV-cytoplasm of Ae.kotschyi and its relatives might be used as the alternative male sterile cytoplasm to replace the T.timopheevii cytoplasm for hybrid wheat breeding.Ikeguchi et al.(1999) proposed that spring-type hybrid wheat may be bred by combination of the 1BL-1RS chromosome and Ae.kotschyi cytoplasm with a new fertility-restorer gene discovered in a wheat variety Kitamiharu 48.Zhang et al.(1996) also proposed the use of CMS-Sv lines as the most effective male sterile cytoplasm.     

桂西北典型喀斯特区生态服务价值的环境响应及其空间尺度特征

Study on ecosystem service values (ESVs) is the bridge of understanding ecosystem and economic decision-making. To investigate the response of ESVs to ambient environment and their spatial scales is an urgent work in the process of ecological restoration and sustainable development in southwest China. Based on the previous research results, the remote images, and weather data of 31 years (1975-2005), the response of ESVs to ambient environment and their spatial scales in a typical karst area of northwest Guangxi, China were evaluated with the method of empirical mode decomposition (EMD) in this paper. The results showed that ESVs had obvious variations with changes in elevation, rocky types and soil types, and had corresponding characteristics of spatial scales. The ESVs in middle and low elevation areas were higher, but they decreased due to human activities such as deforestation. The ESVs in peak-cluster depression areas were lower, and they would increase as influenced by policies such as returning farmland to forestland. The ESVs were influenced by rocky types, showing lower values but an increasing trend in the typical karst regions while higher values but a decreasing trend in non-karst regions. The average ESVs in the west part of the study region, which is not only the main nature reserve for rare wildlife species in Guangxi, but also one of the best preserved natural vegetation regions in China, were more than 15,000 RMB Yuan ha-1,. Comparatively, the ESVs were less than 10,000 Yuan ha-1 in the middle part of the region attributed to low vegetation coverage, serious peak-cluster depression and karst rocky desertification. After about 20 years, that is from 1985 to 2005, the ecosystem conditions had been improved and the ESVs had increased in the middle and eastern part of this study region. On the contrary, in most western part, which was dominated by subtropical evergreen broad-leaved forest, they had been worsened, and the balance values of ESVs were negative. As for the relationships of ESVs with rocky types and soil types, lime soil and red soil were the two main soil types that contributed to ESVs (The total contribution was above 60%, and it is 63.77%,64.37%,64.56%,64.91% respectively in the four years of 1985, 1990, 2000 and 2005.). The variance contribution of intrinsic mode function (IMF) and trend (R) showed that there were obvious special sales for ESVs in this study area, and the main spatial scales were about 2.7 km, 5.5 km, and 11.6 km. The variance contribution ratio was 12.29%, 11.26%, 11.49% respectively, and the trend (R) was high (17.74%). In conclusion, this study indicated that ecosystem conditions in the typical karst area were improved owing to the application of rocky desertification control policies, such as ecological migration and returning farmland to forestland. Besides, the main spatial scales of ESVs in the study area were the interactions of terrain, physiognomy, land use / land cover and human activities.     

Rapid classification of partial waxy wheats using PCR-based markers.

Mutations in the three homeologous waxy loci Wx-A1, Wx-B1, and Wx-D1 of a waxy wheat line have previously been characterized at the molecular level. Using combinations of these mutations, six types of partial waxy wheat plus wild type and waxy wheat (types 1-8) can be produced. Here, we describe primer sets for all three loci that can be used under a single set of conditions, allowing 32 lines to be characterized as types 1-8 in a single PCR run using a 96-well plate. Using multiplex PCR, mutations at the Wx-B1 and Wx-D1 loci can be identified in a single PCR, reducing the number of reactions necessary to identify and select the desired partial waxy wheat line. A single multiplex PCR can be used to detect all three mutations when products are analyzed using capillary electrophoresis on a microchip device. The PCR conditions and primers are effective with a number of cultivars from other countries, indicating that the mutations found at the Wx-A1 and Wx-B1 loci of these cultivars likely have the same origins as the mutations in the corresponding loci of the waxy wheat line used in this study. The PCR selection method described here is an easy and effective alternative to the commonly used SDS-PAGE methods for identification of null alleles.     

Identification of three Wx proteins in wheat (Triticum aestivum L.)

Nullisomic analysis of waxy (Wx) protein of hexaploid wheat (Triticum aestivum L.) cv. “Chinese Spring” using two-dimensional polyacrylamide gel electrophoresis revealed that threeWx loci,Wx-A1, Wx-B1, andWx-D1, located on chromosome arms 7AS, 4AL, and 7DS, produce three distinct Wx subunit groups, subunit group-A (SGA), SGB, and SGD, respectively. SGA has a higher molecular weight and a more basic isoelectric point (pI) than the other two. SGB and SGD have the same molecular weight but a slightly different pI range. Owing to the detection of these three subunit groups, we were able to identify the expression of three waxy genes in wheat endosperm and to find two types of mutants among Japanese wheat cultivars, one lacking SGA and the others SGB. These results suggest the possibility of breeding a waxy wheat.     

小麦蜡质基因座位上SSR的多态性及其与直链淀粉含量的关系

分析了蜡质基因(Wx)3′端的一个简单重复序列(SSR)(AT(AT)n AT)在32份小麦中的多态性及其与直链淀粉含量(AC)的关系。这些材料包括了山东省内的不同小麦品种,其AC含量差别较大。在扩增的小麦品种中均出现两条带,一条204bp(Wx-Dla)位于7DS染色体上,另一条225—346bp(Wx-Ala)位于7AS染色体上。也就是说在扩增的小麦品种中均存在Wx-Ala和Wx- Dla基因,并且较长的片段显示了长度多态性。分析表明AC与7D上这个SSR序列基因的多态性呈正相关,高AC小麦品种扩增的片段较长,电泳迁移率较小;相反,低AC小麦品种扩增的片段较短,迁移率较大,不同长度SSR品种间AC差异显著。虽然目前还不能确定这个SSR序列在直链淀粉合成中的直接功能,但SSR变异与AC之间的相关性可作为分子标记直接用于小麦品种改良。     

利用回交结合Wx基因分子标记培育部分糯性小麦

颗粒结合型淀粉合成酶Ⅰ(GBSSI,Waxy蛋白)是小麦胚乳中直链淀粉合成的关键酶。小麦基因组中存在3个Waxy基因(Wx-A1、Wx-B1、Wx-D1)。在白火麦中Wx-D1位点的突变(Wx-D1b)引起Wx-D蛋白的缺失,导致直链淀粉含量下降,其面粉表现出部分糯性。与目前生产上推广品种相比,白火麦的农艺性状较差,产量非常低。为了培育农艺性状优良的部分糯性小麦,我们将白火麦与具有优良农艺性状的小麦品种PH85-16、济南17和烟农15(轮回亲本)分别进行杂交,后代分别与相应的三个轮回亲本回交五代,在每代中均选择农艺性状与轮回亲本相近并含有Wx-D1b的后代。在第六代自交后选择具有Wx-D1b的纯合体,选出的单株连续自交三代。获得了六个农艺性状与轮回亲本相似的品系,它们均携带纯合的Wx-D1b位点。研究表明采用回交的方法并结合基于Wx基因序列的分子标记技术,是培育优良部分糯性小麦的一种非常有效的方法。本研究培育出的部分糯性小麦品系可以直接用于大田生产。     

Sequences of the Waxy Loci of Wheat: Utility in Analysis of Waxy Proteins and Developing Molecular Markers

The waxy proteins from a number of genetic backgrounds of wheat and its progenitors were analyzed by SDS-PAGE. The deduced amino acid sequences of the waxy proteins of three diploid progenitor species indicated several key amino acid substitutions, which could explain the differences observed in the electrophoretic mobilities of the wheat waxy proteins. A slight difference observed in the apparent molecular weight of the WX-A1 protein of diploid and polyploid wheat was explained by amino acid substitutions or variations in predicted protein structures. Further, twelve different partial genomic clones, representing the individual waxy loci of the various diploid, tetraploid and hexaploid wheats, were isolated and compared. The results indicated significant variations in intron 4 and led to identification of sequences unique to the individual waxy genes and genomes of wheat and its proposed progenitors. The sequence variations observed have a great potential for development as molecular markers for identification of specific waxy loci and study of the various waxy mutants of wheat.     

Detection of DNA sequence polymorphisms among wheat varieties

Summary A DNA marker detection strategy that allows the rapid, efficient resolution of high levels of polymorphism among closely related lines of common wheat (Triticum aestivum) has been developed to circumvent the apparent lack of restriction fragment length polymorphism in many important self-pollinated crop species. The technique of randomly amplified polymorphic DNA (RAPD) was combined with a denaturing gradient gel electrophoresis system (DGGE) to explore DNA sequence polymorphisms among different genotypes of wheat. Of the 65 primer combinations used for the polymerase chain reaction (PCR) amplifications, over 38% of them produced readily detectable and reproducible DNA polymorphisms between a spring wheat line, SO852, and a winter wheat variety, Clark. A high level of polymorphism was observed among a number of commercial varieties and breeding lines of wheat. This procedure was also used to detect polymorphisms in a recombinant inbred population to test the feasibility of its application in genome mapping. This DNA polymorphism detection system provides an opportunity for pedigree analysis and fingerprinting of developed wheat lines as well as construction of a high density genetic map of wheat. Without the need for ³²P and sophisticated DNA extraction procedures, this approach should make it feasible to utilize marker-based selection in a plant breeding program.     

利用回交法与Wx基因分子标记辅助选择培育糯性小麦

以中国春糯性位点全套近等基因系为研究材料,对小麦Wx基因的6个STS标记和1个CAPS标记进行了筛选。改良PCR扩增条件以及产物检测方式后,从这些标记中筛选出3个标记,包括鉴定Wx-A1、Wx-D1位点的2个共显性STS标记和Wx-B1位点的1个显性STS标记,用于本研究中糯性小麦的分子标记辅助育种。在育种过程中,首先配制全糯材料“98Y1441”与推广品种“川育12”的杂交组合,采用籽粒碘染法从其F2种子中选择全糯基因型个体与回交亲本川育12杂交,如此反复自交、回交,历经数代异地加代繁殖得到BC5F2代回交改良群体。利用上述3个分子标记从该群体中筛选出了8种Wx基因型,经卡方检验,其分离比符合3对基因的分离比例,其中基因型为aabbdd的植株有2株,直链淀粉含量分别为1.81%和0.82%,为全糯小麦;基因型为AAbbdd, aabbDD的部分糯性植株各有1株,直链淀粉含量分别为15.24%和17.57%。研究中获得的BC5 F2代群体的农艺性状接近回交亲本,并明显优于全糯材料“98Y1441”,表明采用回交法与Wx基因分子标记辅助选择相结合,有助于培育高产、优质的全糯和部分糯小麦。     

Rapid RAPD screening of plant DNA using dot blot hybridization

Scoring of the results of RAPD analysis using gel electrophoresis imposes a constraint on throughput. To circumvent this barrier, dot-blot hybridization was substituted for electrophoresis. Arbitrarily amplified fragments from barley and wheat genomic DNA were labelled and used as probes for the identification of identical fragments in subsequent amplification reactions. None of the twelve fragments used as probes exhibited significant levels of croos-hybridization to other fragments amplified by the same arbitrary primer. The strength of the hybridization signal facilitates more accurate and more sensitive detection of diagnostic fragments than gel electrophoresis. In addition, the defined spatial orientation (microtitre dish format) of the ± results provide an excellent format for automated data collection. The use of dot blot hybridization to analyse PCR products well decrease the cost and time requirements of marker-assisted selection. This technique will also facilitate the rapid application of PCR-based maps.