苹果腐烂病菌侵染过程中枝条相关病程物质含量的变化

为探究苹果腐烂病的抗病机制,分别用中抗腐烂病种质‘法8’和‘高5’,感病种质‘紫云’和‘红富士’以及高感种质‘嘎拉’和‘金冠’的1年生休眠枝离体接种腐烂病菌株03-8的菌丝,接种后0~7天检测接种部位周围皮层健康组织中病程相关物质多酚、水杨酸和木质素含量变化。结果表明：所有种质接种后1~7天皮层中对香豆酸、肉桂酸和绿原酸含量均显著升高,但仅绿原酸含量与腐烂病的病斑长度显著负相关(r=-0.906)。接菌后皮层组织水杨酸和木质素含量显著增加,水杨酸和木质素含量与病斑长度显著负相关(r=-0.825;r=-0.948)。因此,接种苹果腐烂病菌后皮层水杨酸、木质素和绿原酸含量的显著增加与苹果抗腐烂病相关。     

转Lchi1烟草T_2代对梨腐烂病菌的抗性研究

获得转化梨几丁质酶基因Lchi1的烟草,检测组成型表达Lchi1基因烟草是否能提高对梨腐烂病菌(Valsa ceratosperma)的抗性,从而初步确定Lchi1基因的功能,为进一步开展梨抗腐烂病分子育种工作提供参考。构建梨几丁质酶基因Lchi1的植物表达载体,通过农杆菌介导的叶盘法转化烟草,进而对转化的烟草T2代进行PCR及RT-PCR检测。利用离体叶片接种法,更直观地验证转基因烟草对梨腐烂病菌的抗性。Lchi1基因已经整合到烟草基因组中,并在烟草T2代中有效表达,在接种腐烂病菌后转基因烟草较未转基因烟草的抗腐烂病菌能力明显增强。梨几丁质酶基因Lchi1与腐烂病抗性相关联,可作为今后梨的抗腐烂病分子育种研究重要的基因来源。     

Uniparental inheritance of purple leaf and the associated resistance to leafminer in castor bean

Leafminer ( Liriomyza trifolii Burgess), Diptera Agromyzidae, is one of the insect pests that causes economic damage to castor bean ( Ricinus communis L.) foliage. Green leaf type is a common phenotype in castor bean and highly susceptible to leafminer. The rare purple leaf type germplasm accessions showed stable resistance to leafminer. Studies were carried out to understand the inheritance of purple leaf and the associated leafminer resistance. Direct and reciprocal crosses were made between a purple leaf parent RG1930 and a green leaf parent RG2788. RG1930 is resistant to leafminer while RG2788 is susceptible. Reciprocal differences were noted in segregation pattern of purple leaf colour as well as resistance to leafminer. Purple leaf phenotype was obtained only in purple × green (RG1930 × RG2788) cross where the female parent was a purple leaf phenotype. The reciprocal cross green × purple (RG2788 × RG1930) produced only the green leaf phenotype. Uniparental inheritance was observed for purple leaf phenotype and resistance to leafminer in F₁, F₂, F₃ and backcross generations. Progenies with a dark purple leaf were resistant to leafminer while those with a green leaf were susceptible. Visual association between a purple leaf and resistance to leafminer and their uniparental inheritance were clearly established. The role of heritable epigenetic effects are discussed in expression of purple pigment in offspring.     

Development of high-density interspecific genetic maps for the identification of QTLs conferring resistance to <Emphasis Type="Italic">Valsa ceratosperma</Emphasis> in apple

Valsa canker (Valsa ceratosperma (Tode ex Fr.) Maire) is one of the most destructive fungal diseases of apple, especially in Eastern Asia. In this study, the first high density genetic linkage map of Malus asiatica × Malus domestica was constructed by 640 simple sequence repeats (SSRs) and 490 single nucleotide polymorphisms (SNPs), which spanned 1497.5 cM with an average marker interval of 1.33 cM per marker. Quantitative trait loci (QTLs) for apple’s resistance to V. ceratosperma isolates 03-8 and xc56 were identified using the linkage map. Lesion lengths were used as the phenotypic data for the QTL analysis, which were measured on 1-year-old shoots inoculated with conidia of the two isolates. One QTL for resistance to isolate 03-8 was mapped on LG 16, and one QTL for resistance to isolate xc56 was detected on LG 9. Our research not only promoted the further understanding of the genetic basis of apple’s resistance to Valsa canker but also provided two molecular markers that might be used in future marker-assisted selection for resistance in apple breeding programs.     

Screening methods of apple trees for resistance toValsa canker

Summary Valsa canker is the most serious wood rot disease of apple in Japan. Dormant, excised twig assay in vitro was used to determine resistance to Valsa canker. ‘Fuji’, ‘Starking Delicious’, ‘M.26 EMLA’, and ‘MM.106’ were considered to be highly susceptible, while Malus sieboldii showed moderate resistance. Several cultivars belonging toM. prunifolia, M. baccata, andM. × domestica showed comparative resistance. The phloridzin content in the bark ofMalus species was studied in relation toValsa canker resistance. Phloridzin content ofM. sieboldii was lower than inM. × domestica. Comparatively resistant cultivars had similar or higher phloridzin content compared to susceptible cultivars. No clear correlations were found between resistance and phloridzin contents in the bark ofMalus species.     

生物药剂和化学药剂对苹果树腐烂病菌的抑制效应

为研究不同生物药剂和化学药剂对苹果树腐烂病的影响,分别以多粘类芽孢杆菌、枯草芽孢杆菌、多抗霉素、苦参碱4种生物药剂和戊唑醇、咪鲜胺2种化学药剂为材料,采用培养皿离体培养方式,研究单一药剂和不同药剂组合对苹果腐烂病病原菌丝生长和分生孢子萌发的抑制效应。结果表明：在培养基中添加单一药剂时,多粘类芽孢杆菌、枯草芽孢杆菌浓度为180 mg /L、多抗霉素浓度为225 mg /L、苦参碱浓度为5 mL/L和戊唑醇、咪鲜胺浓度为0.75 mL/L时,抑菌率均可达100%。另外,0.02 mL/L咪鲜胺分别与45 mg/L枯草芽孢杆菌、45 mg/L多粘类芽孢杆菌、0.045 mg/L多抗霉素以及0.02 mL/L戊唑醇与45 mg/L多粘类芽孢杆菌组合,4种组合的菌丝抑菌率与孢子抑制率都达到100%,抑菌效果明显好于其他药剂组合。说明这4种生物药剂和化学药剂组合可作为高效、低毒的混合型药剂来防治苹果树腐烂病的发生。     

苹果腐烂病菌诱导产孢方法

为了研究一种能诱导苹果腐烂病菌快速大量产孢的方法,测试了6种培养基对苹果腐烂病菌的诱导产孢效果。结果发现,苹果腐烂病菌在加蜂蜜水和蛋白胨的带壳大麦上能大量产孢。接种苹果腐烂病菌菌丝的带壳大麦,在黑暗中培养15天后可产生大量黑色子实体。已形成子实体的大麦粒在黑光灯下诱导25天后开始产生分生孢子。每颗大麦粒平均产生11个子实体,其中15%的子实体能溢出分生孢子角。70 g大麦粒上所产生的分生孢子可配制浓度为106个孢子/mL的孢子悬浮液1500~2000 mL。试验还发现,不同的腐烂病菌菌株在同一条件下产孢量存在很大差异。     

Identification of AFLP and RAPD markers linked to anthracnose resistance in grapes and their conversion to SCAR markers

Resistance to anthracnose or black spot ( Elsinoe ampelina ), a serious fungal pathogen in viticulture and table grape production, was investigated on 25 grape cultivars. Bioassays performed with culture filtrates produced by the pathogen revealed 14 resistant genotypes. In most plants resistance originated from Vitis labrucsa but also genotypes with V. rupestris and V. riparia  ×  V. rupestris background showed resistance. Genetic analysis was conducted in F₁, S₁ and BC₁ plants developed from various cultivars. In total, 326 F₁ plants were evaluated, 172 genotypes proofed to be resistant, whereas 154 were susceptible to anthracnose. A Mendelian segregation ratio of 1 : 1 (χ² = 0.30–0.65) indicating that anthracnose resistance is controlled by a single dominant gene. To facilitate the use of marker-assisted selection in grape-breeding PCR-based markers were developed by random amplified polymorphic DNA and amplified fragment length polymorphism in bulk segregant analysis. Finally, OPB 15₁₂₄₇ was developed as a sequence characterized amplified region marker being diagnostic for the locus of resistance to anthracnose in all resistant genotypes tested. Within the 25 grape cultivars OPB 15₁₂₄₇ is diagnostic in the genetic background of both V. labrucsa and V. rupestris and V. riparia  ×  V. rupestris .     

Development of apple powdery mildew on sources of resistance to Podosphaera leucotricha, exposed to an inoculum virulent against the major resistance gene Pl-2

The major resistance gene Pl-2 in apple has recently been shown to be overcome by a virulent population of powdery mildew, Podosphaera leucotricha , in genotypes of the fourth generation after introgression of Pl-2 from crab apple, Malus zumi . Tests of pathogenicity in controlled conditions and observations in an orchard showed that the initial progenitor of Pl-2 , MAL 68/1, was overcome by a virulent inoculum. As a consequence, all selections derived from MAL 68/1 may lose their resistance to powdery mildew, if Pl-2 is used alone. Sources of resistance other than Pl-2 remained efficient to the virulent population: M. hupehensis , M. mandshurica , M. robusta , M. sargentii , M. sieboldii , D12, Mildew Immune Selection and 'White Angel'. The combination of Pl-2 with quantitative resistance genes resulted in a high level of resistance.     

梨树腐烂病在香梨树体上的空间分布特点

为了给香梨树上梨树腐烂病制定有效的调查方法和防治措施提供依据,调查研究了梨树腐烂病在新疆阿克苏地区和库尔勒市香梨树的危害情况及其在树体上的空间分布特点。结果表明：香梨园中梨树腐烂病病株率达100%,其发病程度与梨园的管理水平密切相关。梨树腐烂病在香梨树主干上较少发生,发病率仅1.02%。香梨树不同轮枝（一轮枝、二轮枝、三轮枝）的病斑数、枯枝数差异均极显著;在不同方位（东、西、南、北）上病斑数、枯枝数差异均不显著;在主枝和侧枝上病斑数差异极显著,枯枝数差异不显著;在枝条阴面和阳面病斑数差异极显著。梨树腐烂病主要发生在香梨树的第一轮枝,随着轮枝数的增加,病斑数和枯枝数均显著减少;在香梨树不同方位分布差别不大;其危害部位主要集中在主枝和侧枝,主枝发病较侧枝严重,枝条的阴面较阳面严重。     

Inheritance and gene tagging of male fertility restoration of cytoplasmic-nuclear male-sterile line NJCMS1A in soybean

The F₁, F₂ and F_2:3 of the NJCMS1A × 'Zhongdou 5' cross were used to analyse the inheritance of the male fertility restoration of the cytoplasmic-nuclear male-sterile line NJCMS1A in soybean. The results of genetic analysis showed two pairs of dominant genes conferring the male fertility restoration of NJCMS1A, which further confirmed previous results. The F₂ population from the NJCMS1A × 'Zhongdou 5' cross was used for tagging the restorer genes for NJCMS1A with 664 pairs of simple sequence repeat primers selected randomly from the genetic linkage map of soybean published by Cregan et al. (1999) . Satt626 on linkage group M and Satt300 on linkage group A1 of the integrated linkage map by Song et al. (2004) were found to link to the two restorer genes of NJCMS1A. The maximum-likelihood estimates of the genetic distance between the two markers, Satt626 and Satt300, and the two restorer genes of 'Zhongdou 5' were 9.75 and 11.18 cM, respectively.     

Use of SSR markers to assess sexual vs. apomictic origin and ploidy level of breeding progeny derived from crosses of apple proliferation-resistant Malus sieboldii and its hybrids with Malus × domestica cultivars

To obtain apple rootstocks resistant to apple proliferation and suitable to modern fruit growing, 24 cross-combinations were performed over a 5-year period using Malus sieboldii and its hybrids as donors of the resistance trait and standard apple rootstock Malus  ×  domestica genotypes as donors of agronomic value. Breeding with these genotypes was achieved despite different degrees of apomixis and polyploidy. Sets of five to six locus-specific microsatellite markers were identified for characterizing each progeny. Supported by flow cytometry these markers were applied to infer mode of reproduction, genomic constitution and ploidy level. Microsatellites allele composition identical to the maternal parent was revealed in 1668 of 3032 seedlings indicating seed formation through apomixis. Complete genetic recombination was found in 398 seedlings. The remaining hybrids displayed a higher ploidy than that of the parental plants which was consistent with the fertilization of unreduced egg cells. Thus, for each cross-combination, microsatellite loci were identified which enabled a reliable prediction of the ploidy level. They can now be applied in routine screening to distinguish sexual from apomictic progeny.     

Citrus tristeza virus resistance in a core collection of sour orange based on a diversity study of three germplasm collections using QTL-linked markers

Seven markers linked to QTL involved in CTV accumulation, leafminer resistance and apomictic reproduction were used to characterize 64 sour orange ( Citrus aurantium L.) accessions from three national collections in order to identify a representative core in which the resistance behaviour against two Citrus tristeza virus (CTV) isolates was studied. Different degrees of apomixis were found between the foreign collections. Most of the C. aurantium accessions fall into three main groups based on three multilocus genotypes. The haplotype diversity at three CTV accumulation QTL-linked markers was further studied by sequence analysis of alleles. Genotypic and allelic diversity at one of them, tightly linked to Ctv-R ₂ in Poncirus trifoliata (L.) Raf. , match the plant–CTV interaction types reported among Poncirus and Citrus species. Only those selected accessions from the major group presented CTV resistance during 30 months of continuous growth, but later the resistance broke down in some plants. CTV tolerance appears related to slow growing genotypes. Certain micronutrients: Mn, and B depending on the accession, might play a relevant role in this host–pathogen interaction particularly in alkaline soils.     

Strong evidence for a fire blight resistance gene of Malus robusta located on linkage group 3

Fire blight (FB), caused by the Gram-negative bacterium Erwinia amylovora is a dangerous disease on pome fruit, including apple. The FB-susceptible cultivar 'Idared' was crossed with the resistant wild species clone Malus  ×  robusta 5. A segregating population of 146 progeny has been tested by artificial shoot inoculation for susceptibility to FB. Progeny were infected from 0% to 100% of the shoot length. To identify chromosomal regions or loci responsible for resistance to FB of Malus  ×  robusta 5, a set of microsatellite markers (simple sequence repeat, SSRs) was chosen covering all linkage groups of apple. Up to eight different microsatellites were bulked to one mutliplex PCR using four different labels and a fifth label for a size standard. Fifty-nine microsatellite markers out of 72 SSRs were polymorphic. Fifty-four of 66 loci detected could be mapped and were useful for the detection of related resistant loci. Alleles of microsatellites Hi03d06, CH03g07 and CH03e03 originating from the resistant donor M. robusta were associated with resistance to Erwinia amylovora . Up to eighty percent of the phenotypic variation could be explained by the interval spanned by SSRs CH03g07 and CH03e03, indicating the presence of a major resistance gene. All three microsatellites are located on the distal part of linkage group 3, spanning 15 cM. The SSR marker CH03e03 can be regarded as diagnostic marker for FB resistance. Only seven progeny expressing allele b (184 bp) of CH03e03 showed blighted shoot lengths of more than 30% and only nine progeny lacking allele b showed blighted shoot lengths of <30%. By setting a threshold of 30% shoot necrosis for resistance to FB, the 146 individuals segregate into 71 susceptible and 75 resistant plants, and resistance to FB maps 9 cM away from marker CH03e03.     

Growth and Yield Responses to Plant Density and Stage of Transplanting in Winter Oilseed Rape (Brassica napus L.)

Growth and yield responses to plant density (6.75 × 10⁴, 9.75 × 10⁴ and 12.75 × 10⁴ plants ha^–1) and stage of transplanting (30, 35 and 40 days after sowing) of winter oilseed rape cultivar HO 605 were investigated in two field trials in the 1996/97 and 1997/98 growing seasons at Zhejiang University Farm, Huajiachi Campus, China. Results revealed a progressive decrease in leaf area per plant in response to increasing plant density and delayed transplanting, though leaf area m^–2 and leaf area index were higher in high-density plants. Number of effective branches and pod per branch decreased with increasing plant density and delayed transplanting. There were no significant differences in the mean seed weight among treatments. Although the average number of seeds per pod was significantly lower for high-density plants and delayed transplanting, the economically highest seed yields were realized in relatively high-density plants. Seed oil content was negatively affected by increasing plant density, but no significant differences were observed with delayed transplanting. The highest seed yields of 1730.7 and 1748.1 kg ha^–1 with no significant differences were observed for plant densities of 9.75 × 10⁴ and 12.75 × 10⁴ plants ha^–1, respectively, transplanted at 35 and 30 days after sowing.     

Photosynthetic Activity and Soluble Carbohydrate Content Induced by the Cold Acclimation Affect Frost Tolerance and Resistance to Microdochium nivale of Androgenic Festulolium Genotypes

The aim of the experiment was to verify whether changes in photosynthetic activity measured by means of chlorophyll fluorescence techniques and soluble carbohydrate level induced during pre-hardening and cold acclimation in androgenic forms of Festulolium are related to their winter hardiness and pathogen resistance. The study was performed on six androgenic genotypes of Festulolium, which differ in their degrees of winter hardiness. The chlorophyll fluorescence parameters and sugar content were measured in the control plants, pre-hardened at 12 °C and cold-acclimated at 2 °C. Cold acclimation at 2 °C, preceded by pre-hardening at 12 °C, induced a decline in maximum PSII quantum efficiency ( F _v/ F _m) and in non-photochemical quenching (NPQ). Exposure to 2 °C also resulted in reduced efficiency of energy conversion in photochemical processes ( q _p). The soluble carbohydrate amount increased in the susceptible genotypes and was associated with a decline in NPQ and current quantum yield of PSII (φ_PSII). In resistant plants, a decrease in the sugar content was observed, which could be explained by their consumption in metabolic processes initiated during cold acclimation. The changes in sugar content were associated with a decline in NPQ and q _p.     

苹果腐烂病病原菌的鉴定及拮抗菌筛选

苹果腐烂病是造成华北地区苹果树主要病害之一,爆发时造成苹果树大面积死亡。为开展苹果腐烂病生物防治,本研究从山西阳泉市盂县苹果种植基地的感病植株采集组织样品,利用组织分离的手段获得目标菌株1个,编号为SX1408。采用分子生物学手段,以真菌ITS1和ITS4区通用引物扩增得到ITS区部分序列(Gen Bank登录号KR007599),经Blast和MEGA 5.0聚类分析后,将该菌株鉴定为苹果黑腐皮壳菌(Valsa mali Miyabe et Yamada)。进一步采用平板对峙培养法,从实验室保存的19株供试的生防放线菌中筛选出4株对SX1408有抑菌活性的拮抗菌,其中以BUAS13-3菌株的拮抗效果最佳,抑菌圈直径为2.80 cm,抑制率78%。研究结果将为苹果腐烂病菌的生物防治提供新的生防资源。     

A Quantitative Method to Assess Freezing Resistance in Faba Beans

In order to quantify freezing resistance (R₅₀) of winter faba beans from a limited number of plants, leaves were exposed to artificial freezing stresses with different test temperatures (TT_i). Injury (I_i) was assessed by rating (score: 0—8) and by conductivity measurements. The basic assumption that I_i depends almost linearly on TT_i over a sufficiently broad range with a constant ΔI/ΔTT could be verified and hence a formula is presented to compute freezing resistance R₅₀= (I_i—I₅₀) ×ΔTT/ΔI + TT_i with I₅₀ being a medium injury. This formula allows to quantify resistance by assessing I_i after freezing test(s) with definite TT_i. About 25 leaves are required. Rating appeared preferable to conductivity measurements because of easier and quicker performance and a slightly better discrimination between samples of different resistance. Based on a standardized freezing test, ΔI/ΔTT was determined. Application of the formula with ΔI/ΔTT = 2.5 units/°C for unhardened and hardened leaves, restrictions and experiences are discussed.     

Relationships between seed oil content and fatty acid composition in high stearic acid sunflower

The high stearic acid sunflower mutant CAS-3 is characterized by a low seed oil content, which might represent a constraint for the commercial production of high stearic acid sunflower oil. The objective of the present research was to investigate the relationships between fatty acid profile and seed oil content in CAS-3. Plants of CAS-3 were reciprocally crossed with plants of breeding line ADV-37, with high oil content and standard fatty acid profile. Oil content and fatty acid composition were measured in individual F₂ seeds and F₂ plants (F₃ seeds averaged). Both F₂ seeds and F₂ plants from the cross ADV-37 × CAS-3 had a significantly higher oil content than those from the reciprocal cross, which indicated the existence of cytoplasmic effects in the genetic control of the trait. A consistent negative correlation between oil content and palmitic acid and a positive correlation between oil content and oleic acid were detected both in F₂ seeds and F₂ plants. Conversely, no consistent correlation between oil content and stearic acid was observed, which suggested the feasibility of simultaneous selection for both traits.     

Intra-specific Variation for Salt Tolerance in Linseed (Linum usitatissimum L.)

Thirty-six accessions of linseed ( Linum usitatissimum L.) were screened for salt tolerance at the seedling stage after 2 weeks growth in solution culture salinized with 150mol m⁻³ NaCL Considerable variation in salt tolerance was observed in this set of germplasm. Two salt-tolerant accessions, T-5(69-S9)B and LS-2 and two salt-sensitive accessions P-70 and Aver-Rer-Seic, selected at the seedling stage, were grown to maturity in sand culture salinized with 0 (control), 70, 140, and 210 mol m⁻³ NaCl. Both the salt-tolerant accessions exceeded the two salt-sensitive accessions in biomass production, seed yield, and yield components, but the accessions did not differ significantly in seed oil content in the salt treatments. The tolerant accessions accumulated greater amounts of Na⁺ in the shoots and less in the roots compared with the salt-sensitive accessions. The shoot K⁺, K: Na ratios, and K⁺ versus Na⁺ selectivity were lower in the salt-tolerant accessions compared with the salt-sensitive accessions; the reverse was true for their roots. Contributions of Na⁺ and Ca²⁺ to osmotic adjustment were much higher in the two salt-tolerant accessions than in the salt-sensitive ones.
It has been established that there is a great deal of variation for salt tolerance in linseed. The degree of salt tolerance of this crop does not vary during ontogeny. The physiological basis of salt tolerance of this species appears to be associated with the inclusion of Na⁺ in the shoots.