气候变化对不同水分条件下柿幼树光合作用的影响

以两年生柿树为试材,研究了高浓度CO₂处理（700 μmol·mol^-1）、高温处理（日平均温度高于正常日平均温度约5 ℃）、高温和高浓度CO₂复合处理及对照（温度为外界环境温度,CO₂浓度380 μmol·mol^-1）对不同水分条件下柿树净光合速率(P_n)、蒸腾速率(T_r)、气孔导度(G_s)、水分利用效率(WUE)、叶绿素含量和叶绿素荧光参数F_v/F_m、F_v/F_o的影响.结果表明:高温和高浓度CO₂复合处理使处于各水分条件下的柿树T_r、G_s减小,WUE增大;在高、中水分条件(分别为田间持水量的75%～85%和55%～65%)下高温和高浓度CO₂复合处理使柿树P_n增大,在低水分条件(为田间持水量的35%～45%)下使柿树P_n减小.高浓度CO₂处理使处于各水分条件下的柿树P_n、WUE增大,G_s、T减小.高温及高温和高浓度CO₂复合处理下WUE均受土壤水分状况的影响,并随土壤含水量的升高而升高.与对照相比,高浓度CO₂处理还提高了各水分条件下植株叶片水平的水分利用效率、叶绿素a、叶绿素b、叶绿素（a+b）、类胡萝卜素含量及F_v/F_m和F_v/F_o,缓解了水分胁迫,提高了柿树的抗逆能力.     

Development of microsatellite markers in polyploid persimmon (Diospyros kaki Lf) from an enriched genomic library

The oriental persimmon (Diospyros kaki Lf) is believed to have originated in China with subsequent introduction into Japan and Korea in ancient times. The species was then brought to Europe, Brazil and the USA from Japan in the 19th century. Recent studies highlighted the poor state of identification of cultivars in these countries due to incorrect labelling and presence of synonyms among local varieties. Thus, molecular marker characterization of germplasm resources is of great value for genetic resource preservation and plant breeding of persimmon. Therefore, to identify accessions for further plant breeding and germplasm management, 37 microsatellite loci were developed from a CT/AG‐enriched persimmon genomic library.     

Development of SSR primers using ISSR–PCR in Diospyros kaki Thunb.

Nine polymorphic single sequence repeat (SSR) primers were developed in Japanese persimmon using inter‐SSR (ISSR) suppression polymerase chain reaction (PCR). These primers were tested on 30 individuals from Japan and China. The number of alleles per locus ranged from five to 20. Expected (H_E) and observed (H_O) heterozygosities at each locus ranged from 0.42 to 0.77 and 0.27 to 0.59, respectively. The SSR primers developed herein could be applied to cultivar identification, estimation of genetic diversity and divergence in Diospyros spp.     

应用改良滴冻法超低温保存两种柿属植物的研究

以君迁子（Diospyros lotus L．）和柿（D．kaki Thunb．）组培苗茎尖为试材,对影响超低温保存效果的主要因素,如低温锻炼方式、预培养条件、PVS：（30％甘油＋15％乙二醇＋15％二甲基亚砜＋0．4mol／L蔗糖）处理时间等进行了研究。建立了2种柿属植物的超低温保存程序：（1）切取1cm左右试管苗梢段继代到1／2MS（KNO3和NH4NO3减半）培养基中,交替低温[昼（25±1）℃、夜（4±1）℃]锻炼6周;在含0．5mol／L蔗糖的1／2MS培养基上预培养5d,20℃下装载液（2．0mol／L甘油＋0．4mol／L蔗糖）过渡10min,0℃下PVS2处理1．5h;（2）投入液氮保存;（3）40℃水浴化冻,洗涤5～6次后接种于含1．0mg／LTDZ、0．6g／L可溶性PVP、30g／L蔗糖和7．0g／L琼脂的培养基（作者在优化柿属植物离体培养体系试验中获得）上暗培养1周,转入25℃,1500lx培养室。按照上述程序培养,‘鄂柿1号’、‘湘西甜柿’和君迁子的成活率分别为79．6％、67．4％和60．9％。     

Climatic factors affecting the occurrence of cloudy stain on the fruit skin of ‘Triumph’ Japanese persimmon

Cloudy stain is a physiological disorder of the Japanese persimmon blemishing the skin and causing a great reduction of fruit quality, thus decreasing its commercial value. The stain starts at or just prior to the fruit colour-break stage, coinciding with the presence of cuticular cracks which increase cuticle water permeability and expose subepidermal cells to air and water, causing oxidation of polyphenol substances of large skin areas that turn almost black. The incidence of the disorder examined over four consecutive years in two growing areas of Spain revealed significant differences from year to year, between areas and even the tree face examined. The data consistently showed that cooler groves and fruit located on the northern face of the canopy were more prone to develop the stain. During the 4 years of the study, the average daily minimum temperature of a period of 15–25 days prior to the onset of staining followed the same order as the yearly staining rate, and accumulated hours below 13°C correlated significantly with the percentage of fruit starting the disorder. No relationship was found between the time-course of relative humidity (RH) and the onset of staining, but once the stain started the lower RH resulted in the lower percentage of affected fruit. Dew point, wind speed, rainfall and evapotranspiration did not correlate either with the onset of staining or with its incidence. Rind of cloudy-stained fruit had higher concentration of N, K, Mg and Fe and lower concentration of Ca than that of healthy fruit, but it is not the cause of the disorder. Accordingly, we conclude that cloudy stain of Japanese persimmon starts after a period of low temperature (≤ 13°C) and, afterwards, RH is responsible for its development and severity.     

柿树减数分裂和小孢子形成过程的观察

研究了柿树品种“禅寺丸”（２ｎ＝６ｘ＝９０）的减数分裂和小孢子形成过程。同源染色体配对和分离基本正常,中期Ⅰ构型为４５Ⅱ,但出现似多价体（次级配对）,观察发现了不减数分裂现象。不减数发生的第１次分裂,第２次分裂时可见２个ｎ＝９０的子核。未减数子核在四分体期形成二分体,小孢子期形成大型花粉粒。观察结果提示了柿树多倍体由未减数配子受精产生的可能性。     

柿中具有△^12熊果烯骨架的五环三萜类化合物

柿（Diospyros kaki L.f.）的果实中分离出5个化合物,通过IR、MS、^1HNMR、^13CNMR鉴定,确定为：熊果辛酯（octyl yursolate）,△^5,6－3－（2′－甲在戊酰氧在）－熊果酸环己酯〔△^5,6－3－（2′－methyl pentanoyl）-ursolic acid cyclohexy ester〕,β－香树脂醇己醚（β－amyrin hexyl ether）,△^6,7－香树素（△^6,7－amyin）和熊果甲酯（methyl ursolate）。     

基于叶绿体DNA ndhA和nrDNA ITS序列变异分析栽培柿及其近缘种亲缘关系

以柿属植物（Diospyros spp.）中与柿近缘的8种共30个基因型为试材,进行核糖体DNA(nrDNA)内转录间隔区（ITS）和叶绿体DNA ndhA序列变异分析,并通过软件计算两个序列及合并后的进化模型,依据进化模型采用ML法(maximum likelihood method)分析进化关系。为进一步弄清柿属植物种间亲缘关系和供试柿（Diospyros kaki Thunb.）种内分子差异提供了理论依据。结果表明：（1）ndhA序列长度变异范围在1 492~1 511,14个信息位点;ITS序列长度变异范围在660~761,56个信息位点。ITS、ndhA和ndhA+ITS（ndhA和ITS合并）最适碱基进化模型分别为（TrN+I+G）、（F81+I）和（GTR+I+G）。综合ITS和ndhA序列分析表明：柿与油柿和云南野毛柿亲缘关系最近,与美洲柿和乌柿最远。（2）21份柿品种材料的ITS长度均为730,包括4个变异位点,据此4个变异位点对供试柿种内21个品种进行聚类分析。研究认为,ndhA和ITS能较清楚解释了柿与其近缘种间的亲缘关系,并通过柿品种ITS的差异位点分析鉴别出栽培柿种内的差异。     

Ecology of the methylotrophic bacteria on living leaf surfaces

Abstract Pink-pigmented, facultatively methylotrophic bacteria (PPFMs) are a substantial part of the aerobic, heterotrophic, microflora of young leaf surfaces. They were most numerous on white clover ( Trifolium repens ) leaves in the summer. The pink bacteria averaged 36% of the total heterotrophic count/cm², with a range of 3 to 79%.
The growth of PPFMs on multicarbon compounds is much lower than that of other numerically important leaf heterotrophs. Free methanol of endogenous origin, is present in growing leaves. The availability of methanol at the leaf surface may allow the PPFMs to compete successfully with other heterotrophs that require multicarbon compounds, that are also leached from the growing plant surfaces.
SEM studies showed epiphytic bacteria to be most abundant near the margins of the abaxial surfaces of leaves. Microcolonies of bacteria on leaf surfaces were often covered with a thin layer of material, the source and nature of which has not been determined.     

Rhizobium trifolii mutants deficient in exopolysaccharide production

Spontaneous mutants of Rhizobium trifolii 24AR5 which did not produce exopoly-saccharide were isolated. The non-mucoid mutants formed small white and ineffective nodules on both red and white clover. These nodules contained infection threads, but only a small number of bacteria were released into nodule cells, and bacteroids were rarely observed. The non-mucoid phenotype was not complemented by the symbiotic plasmid (pJB5JI) of Rhizobium leguminosarum.     

Root hair deformation in the white clover/Rhizobium trifolii symbiosis

Rhizobium trifolii most frequently infects its host white clover (Trifolium repens L.) by means of infection threads formed in markedly curled root hairs. Rhizobium infections are classified as either lateral or apical based on whether they originate in the branches or at the apex of the root hairs. A quantitative estimate of lateral and apical infection in the region of the host root (Trifolium repens L. cv. Regal Ladino) that possessed mature and immature root hairs at the time of inoculation with Rhizobium trifolii TAI (CSIRO, Canberra City, Australia) indicated that lateral infection occurred more frequently in the mature root hair region of the root. Apical infections were more common in the immature root hair region. Cell free filtrates collected from R. trifolii cultured in association with the host roots induced branching in white clover root hairs. A partially purified preparation of the branching factor was obtained from freeze-dried filtrates by ethanol extraction and ion exchange chromatography. Preliminary studies on the characteristics of these substances suggest that some are dialyzable and heat stable white others are non-dialyzable and heat labile. The dialyzable, heat-stable compounds contain neutral sugars and range between 1200 to 10000 daltons in size. In roots that were exposed to low concentrations (6–25 μg-ml^?1) of these partially purified deformation factors before inoculation, the developmentally mature root hairs were deformed at the time of inoculation. Nodules appeared in the mature and immature root hair region of these plants at the same time. In plants exposed to water, nodules were observed in the immature root hair region and mature root hair regions 3 and 5 days after inoculation, respectively. Based on these results, we conclude that the nodule development was hastened in the plants exposed to the root hair-deforming substances because the mature root hairs of these plants were made infectible at the time of inoculation by this exposure.     

Incidence and severity of pest and disease damage to white clover foliage at 16 sites in England and Wales

From 1985 to 1987, leaves of white clover ( Trifolium repens ) cv. Grasslands Huia were examined for damage by pests and fungal diseases on seven occasions at up to 16 sites in England and Wales. Pest damage was recorded at all sites on all sampling dates. Over all sampling dates, the mean number of leaves damaged by slugs ranged from 23% to 67%, and that by weevils ( Sitona spp.) ranged from 3% to 62%. Also, up to 30% of leaves were damaged by other, unidentified pests. At individual sites, total pest damage frequently exceeded 90% of leaves. The area of leaf damaged by pests ranged from 2 - 12%. Fungal diseases were recorded in July and September but not in May, and were more prevalent in September 1985 than in September 1986 or 1987. Black blotch ( Cymadothea trifolii ) was the most frequently recorded disease, and the mean number of leaves damaged ranged from 4% to 21%. The mean area of leaves covered by lesions was 1–2%. Infections by viruses were assessed on two occasions, using indicator plants and electron microscopy, and only a very low incidence of arabis mosaic virus and red clover necrotic mosaic virus was recorded.     

合肥市野生地被植物种类构成研究

在野外调查和统计分析的基础上,运用五点随机取样法对合肥市的野生地被资源进行了系统的调查,并根据调查结果,选取发生频率大于30%的野生地被植物作为统计分析的对象。运用Spss10.0软件,对综合值采用组间连接法进行聚类分析,找出合肥市不同类型的野生地被植物的常见种类,并分析其发生特点。结果表明:合肥市野生地被植物种类有119种,涉及43科,105属,其中菊科16种,禾本科10种,蓼科9种,十字花科8种。不同类型的野生地被植物种类构成主要有16种:冬春型野生地被植物4种,夏秋型野生地被植物5种,全光照型野生地被植物4种,蜀山森林生态系统型野生地被植物3种。     

Structure and role in symbiosis of the exoB gene of Rhizobium leguminosarum bv trifolii

The Rhizobium leguminosarum bv trifolii exoB gene has been isolated by heterologous complementation of an exoB mutant of R. meliloti. We have cloned a chromosomal DNA fragment from the R. leguminosarum bv trifolii genome that contains an open reading frame of 981 bp showing 80% identity at the amino acid level to the UDP-glucose 4-epimerase of R. meliloti. This enzyme produces UDP-galactose, the donor of galactosyl residues for the lipid-linked oligosaccharide repeat units of various heteropolysaccharides of rhizobia. An R. leguminosarum bv trifoliiexoB disruption mutant differed from the wild type in the structure of both the acidic exopolysaccharide and the lipopolysaccharide. The acidic exopolysaccharide made by our wild-type strain is similar to the Type 2 exopolysaccharide made by other R. leguminosarum bv trifolii wild types. The exopolysaccharide made by the exoB mutant lacked the galactose residue and the substitutions attached to it. The exoB mutant induced the development of abnormal root nodules and was almost completely unable to invade plant cells. Our results stress the importance of exoB in the Rhizobium-plant interaction. Received: 31 May 1996 / Accepted: 18 December 1996