Isolation, Identification, and Whole-Genome Sequence Analysis of Pantoea Agglomerans Causing Leaf Spot-Hole Disease in Wild Apricot in Xinjiang, China

Wild apricot in Yili wild fruit forest in Xinjiang have been seriously affected by leaf spot-hole disease, with the incidence reaching 100%. To identify the pathogen of apricot perforation in the Yili wild fruit forest, two bacterial strains with strong virulence were obtained by the dilution separation method. The bacterial strains were gram-negative bacteria with yellow colonies, smooth surfaces and neat edges. The results of the pathogenicity test showed that the bacteria could cause symptoms of leaf spot-hole disease in wild apricot, similar to the symptoms in the field, and could cause HR in tobacco. Based on the 16S rDNA gene sequence and multilocus sequence analysis of fusA, gyrB, leuS, pyrG, rpoB and rlpB, combined with the physiological and biochemical characteristics, the isolated strain was identified as Pantoea agglomerans. The pathogen causing bacterial leaf spot-hole disease in wild apricot was determined to be P. agglomerans in the wild fruit forest of Yili, Xinjiang. The whole genome of the pathogen strain GL9-2 was sequenced based on the Illumina HiSeq500 and PacBio RS platforms. The genome size was 4765392 bp, and the G+C value was 55.27%. There was one chromosome and two plasmids in the genome, and 4353 CDs were identified. The annotation results showed that 52 glycoside hydrolase-related genes, 38 bacterial secretory system-related genes and 600 toxin-related genes were predicted.

Elucidating Genetic Diversity in Apricot (Prunus armeniaca L.) Cultivated in the North-Western Himalayan Provinces of India Using SSR Markers

Apricot (Prunus armeniaca L.) is an important temperate fruit crop worldwide. The availability of wild apricot germplasm and its characterization through genomic studies can guide us towards its conservation, increasing productivity and nutritional composition. Therefore, in this study, we carried out the genomic characterization of 50 phenotypically variable accessions by using SSR markers in the erstwhile States of Jammu and Kashmir to reveal genetic variability among accessions and their genetic associations. The genetic parameter results revealed that the number of alleles per locus (Na) ranged from 1 to 6 with a mean Na value of 3.89 and the mean effective number of alleles (Ne) per locus 1.882 with a range of 1.22 to 2. Similarly, the polymorphic information content (PIC) values ranged from 0.464 to 0.104. The observed heterozygosity (Ho) (0.547) was found to have higher than expected heterozygosity (He) (0.453) with average heterozygosity of 0.4483. The dendrogram clustered genotypes into three main clades based on their pedigree. The population structure revealed IV sub-populations with all admixtures except the III sub-population, which was mainly formed of exotic cultivars. The average expected heterozygosity (He) and population differentiation within four sub-populations was 1.78 and 0.04, respectively, and explained 95.0% of the total genetic variance in the population. The results revealed that the SSR marker studies could easily decrypt the genetic variability present within the germplasm, which may form the base for the establishment of good gene banks by reducing redundancy of germplasm, selection of parents for any breeding program.

Phylogeography of Prunus armeniaca L. revealed by chloroplast DNA and nuclear ribosomal sequences

To clarify the phytogeography of Prunus armeniaca L., two chloroplast DNA fragments (trnL-trnF and ycf1) and the nuclear ribosomal DNA internal transcribed spacer (ITS) were employed to assess genetic variation across 12 P. armeniaca populations. The results of cpDNA and ITS sequence data analysis showed a high the level of genetic diversity (cpDNA: HT = 0.499; ITS: HT = 0.876) and a low level of genetic differentiation (cpDNA: FST = 0.1628; ITS: FST = 0.0297) in P. armeniaca. Analysis of molecular variance (AMOVA) revealed that most of the genetic variation in P. armeniaca occurred among individuals within populations. The value of interpopulation differentiation (NST) was significantly higher than the number of substitution types (GST), indicating genealogical structure in P. armeniaca. P. armeniaca shared genotypes with related species and may be associated with them through continuous and extensive gene flow. The haplotypes/genotypes of cultivated apricot populations in Xinjiang, North China, and foreign apricot populations were mixed with large numbers of haplotypes/genotypes of wild apricot populations from the Ili River Valley. The wild apricot populations in the Ili River Valley contained the ancestral haplotypes/genotypes with the highest genetic diversity and were located in an area considered a potential glacial refugium for P. armeniaca. Since population expansion occurred 16.53 kyr ago, the area has provided a suitable climate for the population and protected the genetic diversity of P. armeniaca.

Phylogeography of Prunus Armeniaca L. By Chloroplast DNA and Nuclear Ribosomal Sequences

To clarify the phytogeography of Prunus armeniaca L., two chloroplast DNA fragments (trnL-trnF and ycf1) and the nuclear ribosomal DNA internal transcribed spacer (ITS) were employed to assess the genetic variation across 12 P. armeniaca populations. The results of cpDNA and ITS sequence data analysis showed that the level of genetic diversity in P. armeniaca was high (cpDNA: HT=0.499; ITS: HT=0.876), and the level of genetic differentiation was low (cpDNA: FST=0.1628; ITS: FST=0.0297). An analysis of molecular variance (AMOVA) revealed that most of the genetic variation in P. armeniaca occurred among individuals within populations. The value of interpopulation differentiation (NST) was significantly higher than the number of substitution types (GST), indicating a genealogical structure in P. armeniaca. P. armeniaca shared the same genotypes with related species and may be associated with them through continuous and extensive gene flow. The haplotypes/genotypes of cultivated apricot populations in Xinjiang, North China, and foreign apricot populations were mixed with large numbers of haplotypes/genotypes of wild apricot populations from the Ili River Valley. The wild apricot populations in the Ili River Valley contained the ancestral haplotypes/genotypes with the highest genetic diversity and were located in an area considered a potential glacial refugiume for P. armeniaca. Since population expansion occurred 16.53 kyr ago, the area has provided a suitable climate for the population and protected the genetic diversity of P. armeniaca.

Morphology, DNA Phylogeny, and Pathogenicity of Wilsonomyces carpophilus Isolate Causing Shot-Hole Disease of Prunus divaricata and Prunus armeniaca in Wild-Fruit Forest of Western Tianshan Mountains, China

Prunus divaricata and Prunus armeniaca are important wild fruit trees that grow in part of the Western Tianshan Mountains in Central Asia, and they have been listed as endangered species in China. Shot-hole disease of stone fruits has become a major threat in the wild-fruit forest of the Western Tianshan Mountains. Twenty-five isolates were selected from diseased P. divaricata and P. armeniaca. According to the morphological characteristics of the culture, the 25 isolates were divided into eight morphological groups. Conidia were spindle-shaped, with ovate apical cells and truncated basal cells, with the majority of conidia comprising 3–4 septa, and the conidia had the same shape and color in morphological groups. Based on morphological and cultural characteristics and multilocus analysis using the internal transcribed spacer (ITS) region, partial large subunit (LSU) nuclear ribosomal RNA (nrRNA) gene, and the translation elongation factor 1-alpha (tef1) gene, the fungus was identified as Wilsonomyces carpophilus. The 25 W. carpophilus isolates had high genetic diversity in phylogenetic analysis, and the morphological groups did not correspond to phylogenetic groups. The pathogenicity of all W. carpophilus isolates was confirmed by inoculating healthy P. divaricata and P. armeniaca leaves and fruits. The pathogen was re-isolated from all inoculated tissues, thereby fulfilling Koch’s postulates. There were no significant differences in the pathogenicity of different isolates inoculated on P. armeniaca and P. divaricata leaves (p > 0.05). On fruit, G053 7m3 and G052 5m2 showed significant differences in inoculation on P. armeniaca, and G010 5m2 showed extremely significant differences with G004 7m2 and G004 5m2 on P. divaricata (p < 0.05). This is the first report on shot-hole disease of P. armeniaca (wild apricot) leaves and P. divaricata induced by W. carpophilus in China.

Effect of maturation on wild apricot vermouth of different treatments

Wild apricot vermouths (WAV) of different sugar levels (8, 10 and 12 °Brix), different alcohol levels (15, 17 and 19%) and spice levels (2.5 and 5%) were prepared. The product was matured for six months and evaluated for physico- chemical characteristics at 0, 3 and 6 months of maturation. In general, ethyl alcohol content decreased in wild apricot vermouth of all treatments during maturation for six months, in proportion to their initial values. The decrease in TSS was revealed with the advancement of the storage period of six months. A similar trend was observed for total sugars with the advancement of the ageing period. The amount of reducing sugars, however, increased with the prolongation of the maturation period. The total esters content in WAV increased with the advancement of the ageing period, irrespective of their alcohol content. However, the volatile acidity showed a very little increase during ageing but remained non-significant among the different treatments. Total phenols content in WAV decreased by both the ageing period of six months and an increase in alcohol level. A non-significant increase in the titratable acidity with an ageing period was observed in all the WAV having different alcohol levels. The effect of spice extract levels added in the preparation of WAV showed that with the advancement of storage period total esters increased from 246.8 to 272.8 mg/L and 252.8 to 280.6 mg/L for WAV having 2.5 and 5% spices content, respectively. In brief, an overview of the entire results revealed that there was an interactive effect of alcohol level, the sugar level and the spice extract during maturation of wild apricot vermouth. It can be concluded that the maturation of WAV exerted a favourable effect on physico-chemical properties of wild apricot vermouth and is thus, considered desirable.