Xuesen Chen

Genetic Structure of Malus sieversii Population from Xinjiang, China, Revealed by SSR Markers

One hundred and nine Malus sieversii accessions from four geographical populations located at Kuerdening in Mohe town, Gongliu County, Jiaowutuohai, in Xinyuan County, Daxigou in Houcheng County of Ily State, and Baerluke Mountain in Yumin County of Tacheng State, Xinjiang Uygur Autonomous Region of China were studied by SSR markers. The purpose of the study was to determine the genetic structure and diversity in these eco-geographical populations with eight pair SSR primers of apple. The results indicated that: an average of 16 bands was detected in the four populations. The percentage of polymorphic bands in Gongliu population (89.06%) was the highest in the four populations. The average Neis gene diversity index was 0.257 for all the loci. Totally, 128 polymorphic loci were detected and the percentage of polymorphic loci (P) was 100%, 88.28%, 84.83%, 87.50%, and 87.12%, respectively, at the species level and Gongliu, Xinyuan, Huocheng, and Yumin population levels. The Neis gene diversity index (H = 0.2619) and Shannons information index (I = 0.4082) in the species level were higher than in the population level. The Neis gene diversity index and Shannons information index in the four populations were Gongliu > Huocheng > Xinyuan > Yumin. Gongliu population and Xinyuan population were the highest in genetic identity and the closest in genetic distance. Gene flow between the populations was 7.265 based on genetic differentiation coefficient (G(ST) = 0.064). The UPGMA cluster analysis indicated that the genetic relationships between the Gongliu and Xinyuan population were the closest, and the Yumin population were the farthest with the other three populations. The UPGMA cluster analysis indicated that the four geographical populations located in Gongliu, Xinyuan, Huocheng, and Yumin were relatively independent populations. Concurrently, there was also mild gene exchange between the populations. On the basis of the study of population genetic structure and the highest genetic diversity, Gongliu population should be given a high priority consideration in Malus sieversii populations in situ germplasm conservation.

Identification of self-incompatibility (S-) genotypes of Chinese apricot cultivars

The S-genotypes of 16 apricot (Prunus armeniaca L.) cultivars native to China were determined by the S-allele PCR approach and the results were confirmed by cross-pollination tests among these cultivars. Primer combination EM-PC2consFDxa0+xa0EM-PC3consR, based on the conserved regions C2 and C3 of RosaceousS-RNase genes, was the most useful primer combination for identifying Chinese apricot S-alleles. Twelve S-RNase alleles were identified using this primer combination, and they were defined as follows: S9 was 657xa0bp, S10 was 266xa0bp, S11 was 464xa0bp, S12 was 360xa0bp, S13 was 401xa0bp, S14 was 492xa0bp, S15 was 469xa0bp, S16 was 481xa0bp, S17 was 487xa0bp, S18 was 1337xa0bp, S19 was 546xa0bp and S20 was 1934xa0bp. S11–S20 were new S-RNase genes deposited in GenBank under accession numbers DQ868316, DQ870628-DQ870634, EF133689 and EF160078, respectively. Our findings contribute to a more efficient breeding program of Chinese apricot and further studies on the S-RNase genes.

Population genetic structure in apricot (Prunus armeniaca L.) cultivars revealed by fluorescent-AFLP markers in southern Xinjiang, China.

Population-wide genetic structure was studied using fluorescent-AFLP markers on 85 apricot (Prunus armeniaca L.) cultivars collected from Kuche, Kashi, Hetian in the Tarim Basin, southern Xinjiang Uygur Autonomous Region of China. The purpose of this study was to determine the genetic structure and genotypic diversity among the different eco-geographical populations. Based on the results from this study, 8 pairs of fluorescent-AFLP primers showed clear electrophoregram and high polymorphism amongst the 64 pairs of EcoR|/Mse|(Mse|--a FAM fluorescent marked primer) primers screened. There was a significant polymorphic difference for the same primer pair in different populations and for the same population with different primer pairs. The percentage of polymorphic loci (P) at species level was higher than Kuche, Hetian, Kashi population levels, respectively. The Neis gene diversity index (H) and Shannons information index (I) at species level were higher than those of Kuche, Hetian, and Kashi at population level, respectively. H and I of Kuche population were the highest amongst the three populations. Apricot population genetic diversity was found mainly within the population. Genetic differentiation coefficient between populations (G(ST)) was 0.0882. Gene flow Nm between the populations was 5.1689. Population genetic identity was between 0.9772-0.9811 and genetic distance was between 0.0191-0.0232. These results further indicated that the similarity between populations was higher and the genetic distance between populations was smaller. The UPGMA cluster analysis indicates that the geographical populations at Kuche, Kashi, Hetian were relatively independent Mendelian populations. Concurrently, there was also partial gene exchange between the populations. All the evidences indicated that the genetic diversity in Kuche population was the highest, suggesting that it could be a transition population from wild apricot to cultivated apricot. There were abundant genetic diversities in apricot cultivar populations in southern Xinjiang, China, which provide promising germplasm for further breeding and theoretical basis for biodiversity conservation and utilization for apricot population in this area.

Detection and Transcript Expression of S-RNase Gene Associated with Self-Incompatibility in Apricot (Prunus armeniaca L.)

The identity and expression of S-RNase genotypes in the self-compatible (SC) apricot cultivar ‘Katy’ and the self-incompatible (SI) cultivar ‘Xinshiji’ were examined. We used allele specific polymerase chain reaction (AS-PCR) and designated the alleles in ‘Katy’ and ‘Xinshiji’ as S8Sc and S9S10, respectively. The S-RNase gene was expressed in style at the balloon stage in both genotypes. Using real-time fluorescence quantification RT-PCR technology (FQRT-PCR), spatio-temporal expression patterns of S-RNase gene between ‘Katy’ and ‘Xinshiji’ were compared. The results revealed that the expression of the S-RNase gene in ‘Katy’ and ‘Xinshiji’ were different. The transcript abundance was distinctly diverse at the key stage (i.e., at 24xa0h after self-pollination) in both genotypes, and was greater in ‘Xinshiji’ (SI) than ‘Katy’ (SC). In addition, the abundance of the S-RNase transcript was higher in upper-half of style than in the lower-half of style or in the ovary. In the SI cultivar ‘Xinshiji’, the expression of S-RNase reminded a relatively high level after cross-pollination, but it dropped continuously after self-pollination and un-pollination.

Genetic Diversity of Volatile Components in Xinjiang Wild Apple (Malus sieversii)

To evaluate genetic relationships using qualitative and/or quantitative differentiation of volatile components in Xinjiang Wild Apple (Malus sieversii (Lebed.) Roem.) and to acquire basic data for the conservation and utilization of the species, aroma components in ripe fruit of M. sieversii obtained from 30 seedlings at Mohe, Gongliu County, Xinjiang Autonomic Region, China, and in ripe fruit of 4 M. pumila cultivars (Ralls, Delicious, Golden Delicious, and Fuji) were analyzed using head space-solid phase microextraction and gas chromatography-mass spectrometry. The results indicated that the values of similarity coefficient concerning volatile types between the two species were in accordance with the evolution of M. pumila cultivars (forms), and that M. sieversii seedlings showed considerable genetic variations in these aspects: the total content of volatile components, the classes and contents of each compound classes, the segregation ratio, and content of main components. The results showed significant difference among seedlings and wide genetic diversity within the populations. Comparison of the volatile components in M. sieversii with those in M. pumila cultivars showed that the common compounds whose number were larger than five with the contents over 0.04 mg/L simultaneously between M. sieversii and M. pumila cultivars belonged to esters, alcohols, aldehydes or ketones. This suggests fundamental identity in main volatile components of M. sieversii and M. pumila cultivars. The results above sustained the conclusion M. sieversii is probably the ancestor of M. pumila. However, there were 48 compounds present in M. pumila that were not detected in M. sieversii, including 6 character impact components (i.e., propyl acetate, (Z)-3-hexenal, 2-methyl-1-butanol acetate, pentyl acetate, 3-furanmethanol, and benzene acetaldehyde). This suggested that in the domestication of M. pumila, introgression of other apple species, except for M. sieversii, by interspecies hybridization was possible. There were 177 compounds in total belonging to 11 classes detected in 30 M. sieversii seedlings, including esters, alcohols, ketones, aldehydes, acids, benzene ramifications, terpenes, heterocycles, hydrocarbon derivates, acetals, and lactones. Among them, acetals and lactones were not detected in M. pumila cultivars, 90 compounds were unique to M. sieversii, and 7 components (1-butanol, ethyl butanoate, 1-hexanol, ethyl hexanoate, 3-octen-1-ol, ethyl octanoate, and damascenone) belonged to character impact odors. Thus, the potential of M. sieversii in utilization conservation is enormous as a rare germplasm on genetic improvement of M. pumila cultivars.

Primary molecular features of self-incompatible and self-compatible F1 seedling from apricot (Prunus armeniaca L.) Katy × Xinshiji

Expression of the S-RNase genes in the self-compatible (SC) apricot (Prunus armeniaca L.) cultivar Katy, the self-incompatible (SI) cultivar Xinshiji and their F1 seedling was examined in this study. Three S-genotypes, S9Sc (Sc, self-compatibility S-gene absent from the style), S8S9, and S8S10, were obtained. Seedlings with S-RNase that migrated as a single band in gel electrophoresis were SC, despite high transcript abundance, and those with S-RNase that migrated as two bands were SI with high transcript abundance or SC with low transcript expression. S8-RNase was induced in SI cultivars only 24xa0h after self-pollination, indicating post-transcriptional regulation of S8-RNase in SI apricots. A Proteomic study showed that 35 protein spots were synthesized differently between SC and SI pistils. Fifteen of the 35 protein spots were identified; nine proteins, including receptor protein kinase-like protein, reversibly glycosylated polypeptide-2, and isoflavone reductase-like protein, were detected only in the SC pistils; while nine proteins, including actin 7, a putative serine/threonine kinase, and S-RNase, were detected only in the SI pistils. A mitochondrial NAD-dependent malate dehydrogenase and a probable elongation factor G were up-regulated, while heat shock cognate 70 was down-regulated in the SC pistils compared to those in the SI pistils. The results suggest that the proteins responsible for self-compatibility and self-incompatibility may be different.

Inheritance and Correlation of Self-Compatibility and other Yield Components in the Apricot Hybrid F1 Populations

SummaryTo provide genetic basis for apricot (Prunus armeniaca Lam.) breeding, inheritance and correlation of yield components including self-compatibility, self-pollinated fruiting rate, fertile flower rate, average fruit weight and fruit number per plant were studied with 5-year-old seedlings of apricot F1 hybrids from ‘Katy’ × (‘Xinshiji’, ‘Katy’ × (‘Hongfeng’ and ‘Katy’ × (‘Taianshuixing’, respectively. Using the criteria that cultivars with self-pollinated fruiting rate ≥6% were self-compatible(SC), we found that the ratios of self-compatible (SC) to self-incompatible (SI) individuals were 27:25, 9:12 and 15:19 in the above three families, respectively, and conformed to the ratio of 1:1 segregation by χ2 test, indicating that the S-locus of ‘Katy’ was heterozygous and self-compatibility was dominant to self-incompatibility. Twenty-seven seedlings from the F1 population of ‘Katy’ × ‘Xinshiji’ were chosen for S-allele-specific PCR. As a result, four S-genotypes with the ratio of 10:6:4:7 were obtained, which was linear to the 1:1:1:1 ratio by χ2 test. Great differences in self-compatibility degree were observed among seedlings even with the same S-genotype. In the F1 populations, a very extensive segregation in self-pollinated fruiting rate, fertile flower rate and average fruit weight was observed, and average values of these traits were lower than that of mid-parent. Therefore, these traits were confirmed to be quantitative. However, significant differences were found in broad heritability (Hb2) of following three characters: the Hb2 of self-pollinated fruiting rate (87.1% – 91.4%) was the greatest, with the variation mainly resulted from inheritance; fertile flower rate (36.8% – 49.1%) was the least and its variation was mainly caused by environmental factors. In addition, self-pollinated fruiting rate and fertile flower rate had very significantly positive correlations with single plant yield, so both may play important roles in the determination of single plant yield. In contrast, correlation between yield and average fruit weight was not significant.

Method of Constructing Core Collection for Malus sieversii in Xinjiang, China Using Molecular Markers

Abstract The method for constructing core collection of Malus sieversii based on molecular marker data was proposed. According to 128 SSR allele of 109 M. sieversii , an allele preferred sampling strategy was used to construct M. sieversii core collection, using the UPGMA (unweighted pair-group average method) cluster method according to Nei & Li, SM, and Jaccard genetic distances, by stepwise clustering, and compared with the random sampling strategy. The number of lost allele and t -test of Neis gene diversity and Shannons information index were used to evaluate the representative core collections. The results showed that compared with the random sampling strategy, allele preferred sampling strategy could construct more representative core collections. SM, Jaccard, and Nei & Li genetic distances had no significant difference for construction of M. sieversii core collection. SRAP (sequence-related amplified polymorphism) data and morphological data showed that allele preferred sampling strategy was a good sampling strategy for constructing core collection of M. sieversii . Allele preferred sampling strategy combined with SM, Jaccard, and Nei & Li genetic distances using stepwise clustering was the suitable method for constructing M. sieversii core collection.

Interspecific hybridization of Prunus persica with P. armeniaca and P. salicina using embryo rescue

Embryo rescue technique was used successfully to produce interspecific hybrids by crossing peach (P. persica) as a female parent with apricot (P. armeniaca) and plum (P. salicica). In those crosses that had ‘Yuhualu’ or ‘Zhonghuashoutao’ as female parents, hybrid embryos aborted from the 7th or 8th week after pollination mainly due to post-pollination incompatibility. An embryo rescue protocol was established to rescue such embryos and recover hybrid plants. Modified half-strength MS medium containing 4xa0mgxa0l−1 6-BA and 0.5xa0mgxa0l−1 IBA produced up to 90% germination in the embryos. Modified MS medium with 1.0xa0mgxa0l−1 6-BA and 1.0xa0mgxa0l−1 IBA gave the highest bud induction and multiplication whereas modified MS medium containing 0.5xa0mgxa0l−1 IAA and 0.2xa0mgxa0l−1 NAA gave the best rooting percentage. All the hybrids obtained using this embryo rescue technique were verified using simple sequence repeat (SSR) markers. A series of pollen treatments were carried out to partially overcome pre-pollination incompatibility, and it was found accidentally that pollen treatment with electrostatic field not only improved pollen germination but also increased the multiplication coefficient of embryo-induced shoots.

Comparison of Headspace Solid-Phase Microextraction with Simultaneous Steam Distillation Extraction for the Analysis of the Volatile Constituents in Chinese Apricot

Abstract Volatile constituents in fully mature fruits of apricot (Prunus armeniaca L.) cultivar Xinshiji were extracted using headspace solid-phase microextraction (HS-SPME) and simultaneous steam distillation extraction (SSDE) and then analyzed using capillary gas chromatography and gas chromatography-mass spectrometry. A total of 70 components were identified by HS-SPME, including 20 esters, 19 hydrocarbons, 5 alcohols, 5 ketones, 4 acids, 4 lactones, 3 aldehydes, and 10 miscellaneous components, with the esters being the dominant constituent. On the basis of the odor unit values, it is believed that the following compounds probably contributed to the fresh apricot odor: hexyl acetate, β-ionone, butyl acetate, (E)-2-hexcnal, linalool, limonene, γ-decalactone, and hexanal. A total of 49 components were also detected by SSDE, including 13 hydrocarbons, 9 alcohols, 7 aldehydes, 9 esters, 4 ketones, 4 lactones, 2 acids, and 1 miscellaneous component, of which the monoterpene alcohols were the dominant constituents. It could be judged from the odor unit values that the following compounds were the major contributors to boiled apricot aroma: β-ionone, linalool, hexyl acetate, γ-dodecalactone, γ-decalactone, (E)-2-hexenal, hexanal, γ-octalactone, phenylacetaldehyde, butyl acetate, limonene, α-terpineol, and δ-decalactone. The results show that HS-SPME is a simple, rapid, and solvent-free method, which is an alternative to the classical SSDE.