Investigation and genetic mapping of a Glomerella leaf spot resistance locus in apple

Apple Glomerella leaf spot (GLS) is a severe fungal disease that damages apple leaves during the summer in China. Breeding new apple varieties that are resistant to the disease is considered the best way of controlling GLS. Fine mapping and tightly linked marker are critically essential for the preselection of resistant seedlings. In this study, a population of 207 F₁ individuals derived from a cross between ‘Golden Delicious’ and ‘Fuji’ was used to construct a fine simple sequence repeat (SSR)‐based genetic linkage map. The position of R_gls, a locus responsible for resistance to GLS, was identified on apple linkage group (LG) 15 using SSR markers CH05g05 and CH01d08, which was adapted from a published set of 300 SSR markers that were developed using the bulked segregant analysis (BSA) method. These two SSR markers flanked the gene, and its recombination rate was 8.7% and 23.2%, respectively. A total of 276 newly developed SSR markers around the target region and designed from the genome apple assembly contig of LG15 were screened. Only nine of these were determined to be linked to the R_gls locus. Thus, a total of 11 SSR markers were in linkage with R_gls, and mapped at distances ranging from 0.5 to 33.8 cM. The closest marker to the R_gls locus was S0405127, which showed a genetic distance of approximately 0.5 cM. The first mapping of the gene R_gls was constructed, and the locations of the 11 effective primers in the ‘Golden Delicious’ apple genome sequence were anchored. This result facilitates better understanding of the molecular mechanisms underlying the trait of resistance to GLS and could be used in improving the breeding efficiency of GLS‐resistant apple varieties.     

胡椒园间作槟榔体系小气候对胡椒产量的影响

为确定胡椒园间作槟榔体系影响胡椒产量的关键小气候因子,在海南胡椒园间作槟榔优势区开展4种间作模式与单作胡椒园小气候因子对产量影响的为期4 a的试验,研究不同间作密度下,对胡椒产量有主要贡献的灌浆期的叶片光合作用参数、每日最高和最低气温、不同深度土层土壤温度和含水量,分析胡椒园间作槟榔体系小气候因子对胡椒产量的影响。结果表明:合理间作可以提高胡椒产量,其中间作槟榔株数为765株/hm2的T2处理可以显著提高胡椒产量38.71%,为增产效应最大的处理。分析T2处理增产的原因,发现光强影响的关键光合参数气孔导度对产量有极显著正效应,光合有效辐射、光合速率对产量有显著正效应,T2处理显著提高了这三个光合参数;高温干旱的灌浆期胡椒园日最高温和升温幅度与产量负相关,T2处理显著降低了这两个参数,使胡椒产量增加;表层土壤温度与产量呈负相关关系,T2处理显著降低了表层土壤温度,对胡椒根系有保护作用。综合分析,发现光合有效辐射、日最高温、升温幅度和表层土壤温度是影响胡椒产量的关键小气候因子,所以,这几种小气候因子是用来确定间作模式是否合理的关键参数。     

黑蒜非发酵法制备工艺与抗氧化活性研究

为了提高黑蒜的生产效率、降低黑蒜的生产成本,将蒸制工艺与烘制工艺相结合,设计并优化了一种非发酵黑蒜制备工艺。研究结果表明,非发酵黑蒜的最佳制备工艺参数为:蒸制温度127℃、蒸制时间70 min、烘制温度95℃、烘制时间6.5 h,在此工艺条件下制得的黑蒜总酚质量比为11.15 mg/g,含水率为27.41%,还原糖和总酸质量比分别为7.87 g/(100 g)和36.09 g/kg。在0.06~0.18 mg/m L质量浓度范围内,黑蒜的还原能力是维生素C的1.4倍以上,说明黑蒜具有很强的抗氧化活性。与发酵工艺相比,总酚等活性成分含量相近,但极显著地缩短了黑蒜的制备时间、提高了生产效率、降低能耗,为功能性蒜制品的开发利用提供了技术基础。     

Multilocus sequence typing analysis of Italian Xanthomonas campestris pv. campestris strains suggests the evolution of local endemic populations of the pathogen and does not correlate with race distribution

Xanthomonas campestris pv. campestris (Xcc) is the causal agent of black rot in Brassicaceae. It is widespread in Italy and severe outbreaks occur under conditions that favour disease development. In this study a multilocus sequence typing approach (MLST) based on the partial sequence of seven loci was applied to a selection of strains representative of the main areas of cultivation and hosts. The aim was to investigate whether the long tradition of brassica crops in Italy has influenced the evolution of different Xcc populations. All loci were polymorphic; 14 allelic profiles were identified of which 13 were unique to Italian strains. Based on the seven loci, the most common genotype within the Italian Xcc strains (AP1) was also the most representative genotype found in worldwide Xcc strains. This genotype was included in a new clonal complex in addition to three other clonal complexes already identified in Xcc populations. The phylogenetic reconstruction using a concatenated dataset of four conserved protein-coding genes, dnaK, fuyA, gyrB and rpoD, showed that the Italian strains belonged to two genetic groups. Physiological races were also investigated for the first time in Italy. The race structure of Xcc was determined by inoculating eight differential Brassica lines belonging to five species and showed that, in Italy, race 4 is the most widespread, followed by races 1 and 6. No correlation was found between allelic profiles, host of isolation, geographical origin and races, although a prevalent race was identified within the same clonal complex.     

Genetic diversity of the pine pathogen Lecanosticta acicola in Slovenia and Croatia

Brown spot needle blight (BSNB), a disease of pine trees caused by the fungus Lecanosticta acicola, has been known in Slovenia since 2008 and in Croatia since 1975. Recent outbreaks in Slovenia prompted this study to compare L. acicola populations in these two neighbouring European countries. Sixty-nine isolates collected from three pine species (Pinus mugo, P. halepensis and P. nigra) were used to determine the phylogenetic relationships, genetic structure, and reproductive strategy of the pathogen. EF1-α sequences showed that Slovenian and Croatian isolates share a common ancestry with individuals from central and northern Europe. Population structure analysis revealed four distinct population clusters of L. acicola in these two countries, generally corresponding to their respective geographic location and host. An unequal ratio of mating types and a low overall genetic diversity in the population indicated a strong influence of asexual reproduction. Although some of the oldest recorded European occurrences of BSNB are from Croatia, this study provided no evidence that the population studied in Croatia was the source of the sampled outbreaks in Slovenia. Recent outbreaks of L. acicola in Slovenia are most likely due to introductions from other, yet to be identified, sources.     

Evaluating diverse systems of tobacco genetic resistance to Phytophthora nicotianae in Yunnan,China

Black shank, caused by the soilborne pathogen Phytophthora nicotianae, is one of the most devastating diseases affecting tobacco production in China. The most effective strategy for reducing economic loss from this pathogen is development and use of resistant tobacco varieties. Multiple sources and systems of resistance have been developed in the Western Hemisphere; however, populations of P. nicotianae are variable around the world, including the predominance of different races. Different P. nicotianae isolates may react differently on tobacco plants with different systems of resistance, a possibility that could complicate the breeding of cultivars with resistance that is effective in different tobacco production regions worldwide. The objective of this research was to evaluate an array of tobacco germplasm possessing different systems of genetic resistance to black shank disease in tobacco-growing regions of Yunnan, China. Resistance types included simply inherited resistance mechanisms introgressed from wild Nicotiana relatives and polygenic partial resistance systems of N. tabacum origin. The loci of Wz exhibited high level resistance to black shank in the five diverse disease environments in Yunnan, China. K326 Php/−Wz/− genotype and Beinhart 1000 exhibited the greatest levels of resistance in both 2015 and 2016. Field observed results for 13 tobacco genotypes were highly correlated with those tested in growth chamber evaluation. These findings suggest that both Wz− and Beinhart 1000-mediated resistance have important commercial value in flue-cured tobacco breeding programmes in China. Cultivars developed for black shank resistance in China may also have utility in other tobacco-growing areas.     

甜瓜黄斑病毒在海南黄瓜上的发生分布及序列分析

 应用特异性引物MYSV-L-F和MYSV-L-R对海南5个市县采集的150份疑似感染病毒病的黄瓜样品进行RT-PCR检测,在三亚、乐东和东方等3个市县的18份样品中检测到甜瓜黄斑病毒(Melon yellow spot virus, MYSV)。选取采自三亚的分离物C29(MYSV-Hainan)进行MYSV全基因组克隆及序列分析,结果表明:该分离物的L RNA、M RNA和S RNA基因组全长分别为8 918 nt、4 815 nt和3 257 nt,与已知MYSV分离物的核苷酸相似性分别为97.4%~97.7%、96.2%~97.9%和94.8%~99.8%,系统进化关系分析与相似性分析一致,序列相似值越高,系统进化关系越近。     

贵州省白及叶褐斑病病原菌鉴定及其生物学特性

为明确贵州省白及叶褐斑病病原菌及其生物学特性,采用组织分离法和离体接种法分别对其病原菌进行分离和致病性测定,利用形态学及ITS、EF-1α、β-tubulin基因序列分析对病原菌的分类进行鉴定,并对病原菌的生物学特性进行研究。结果表明:分离得到的白及叶褐斑病病原菌菌株编号为HGUP_(17)355;该菌菌丝白色、羊绒状,菌落中心隆起呈淡黄色、背面枫叶红色,大型分生孢子窄镰刀形,3～8个分隔,未见厚垣孢子,小型分生孢子卵圆形,1～2个分隔;在以ITS、EF-1α、β-tubulin三个基因序列构建的系统发育树中,菌株HGUP_(17)355与燕麦镰刀菌Fusarium avenaceum聚于一支,且支持率达100%,结合形态特征与分子生物学将其鉴定为燕麦镰刀菌。该菌菌丝最适生长温度为20℃,最适生长pH为6,在燕麦片琼脂培养基上生长最快,以麦芽糖为碳源、以L-苯丙氨酸为氮源时利用率最高,致死温度为45℃,对光照不敏感。