Twenty-four representative melon varieties and six parental cultivars were examined in this study. Among 159 pairs of simple sequence repeat (SSR) primers, 18 SSR core primers with rich polymorphic information, a large number of genotypes, and the ability to distinguish different melon varieties were selected. A total of 113 genotypes were detected among the 30 experimental materials, with an average of 6.28 genotypes for each pair of primers. The polymorphic information content was on average 0.6807, ranging from 0.5618 to 0.7885. Specific bands of the primers for the 30 experimental materials were analyzed, and by combining different primer loci, all 30 varieties were identified. Unique barcodes for molecular identity cards for the 30 experimental materials were established using the fingerprints formed with this SSR marker system. Each variety has a unique identity card that can be applied for the registration of the newly bred varieties, the protection of breeders’ rights, and the authenticity of breeds after promulgation of the new Seed Law of the People’s Republic of China.
Li Li, Xiulan Xu, Ping Wu, Guo Zhang and Xiaobing Zhang
Li-ping Chen, Yan-ju Wang and Man Zhao
In this study, in vitro induction of tetraploid Lychnis senno Siebold et Zucc. and its cytological and morphological characterization were conducted. For polyploid induction, nodal segments with axillary buds from in vitro grown plants were kept for 3 days in MS (Murashige and Skoog, 1962) liquid or solid media added with a series of concentrations of colchicine. Out of total 588 recovered plants, 15 tetraploids and 6 mixoploids determined by flow cytometry analysis were obtained. The tetraploid contained 48 chromosomes, twice the normal diploid number of 24, as observed under light microscope. The tetraploid plants exhibited much larger but less stomata than diploid plants. Moreover, significant differences in stem height and leaf size between the diploid and tetraploid plants were noted. The tetraploid plants were more compact than diploids.
Li Li, Ling Liu, Deshuang Zhang, Ping Wu, Fenglan Zhang and Xiulan Xu
Thirteen Chinese cabbage (Brassica rapa) hybrid cultivars and 26 parental inbred lines were used as experimental materials to screen for primers producing hybrid and parental complementary bands and for primers with high polymorphism information contents and low genotype frequencies. A total of 18 pairs of core primers were designed to identify the purity of Chinese cabbage. There was no significant difference in the purity percentage measured between different loci of the same strain. The fingerprint obtained by the amplification of each locus could be used to identify purity to obtain an authentic purity percentage. Curve mapping and significance analyses were conducted using the purity percentage of eight different seed samples and confirmed a sampling seed number of 96. The results of the purity test were verified by comparison with the grow-out test (GOT) using molecular markers. In conclusion, the simple sequence repeat (SSR) detection system could be used for the rapid identification of the purity of the tested Chinese cabbage hybrids.
Dong Liu, Ping Li, Jiulong Hu, Kunyuan Li, Zhenyu Zhao, Weiyan Wang, Jinyuan Zhang, Xu Ding and Zhimou Gao
To explore genetic differentiation and the genetic relationships of Phytophthora sojae in Anhui Province, the inter-simple sequence repeat (ISSR) technique was used to analyze the genetic diversity of P. sojae. One hundred and sixty ISSR fragments were observed, including 129 (80.6%) polymorphic bands. This suggested that abundant genetic diversity existed among P. sojae in Anhui Province. The pairwise genetic similarity coefficients among the 62 strains ranged from 0.72 to 0.96, with a mean value of 0.85, indicating that there was a high level of genetic variation. Phytophthora sojae strains were divided into five clusters based on neighbor-joining (NJ) analysis, and the clustering was not related to geographic source, according to Mantel’s test (r = 0.3938). In addition, the clustering of strains from the same geographical source had little relationship to the year of collection. Analysis of molecular variance (AMOVA) showed that 16.65% of the genetic variation was derived from the collection area and 83.35% of the genetic variation was within-population variation. The genetic flow between different geographical sources ranged from 0.623 to 2.773, with a mean of 1.325, suggesting that gene exchange was frequent. Genetic distance and the genetic differentiation coefficient were not related to spatial distance.
Ping Li, Dong Liu, Min Guo, Yuemin Pan, Fangxin Chen, Huajian Zhang and Zhimou Gao
Sexual reproduction in the plant parasite Phytophthora capsici Leonian requires the interaction of two distinct mating types, A1 and A2. Co-occurrence of these mating types can enhance the genetic diversity of P. capsici and alter its virulence or resistance characteristics. Using an intersimple sequence repeat (ISSR) screen of microsatellite diversity, we identified, cloned, and sequenced a novel 1121-base pair (bp) fragment specific to the A1 mating type of P. capsici. Primers Pcap-1 and Pcap-2 were designed from this DNA fragment to specifically detect the A1 mating type. Polymerase chain reaction (PCR) using these primers amplified an expected 997-bp fragment from known A1 mating types, but yielded a 508-bp fragment from known A2 mating types. This PCR-based assay could be adapted to accurately and rapidly detect the co-occurrence of A1 and A2 P. capsici mating types from field material.
Jing Tian, Li-Ping Wang, Yan-Juan Yang, Jin Sun and Shi-Rong Guo
Heat tolerance is considered to be an essential feature for cucumber (Cucumis sativus) production, and it has been suggested that higher antioxidant ability could prevent the oxidative damage in plants caused by high-temperature stress. We aimed to investigate whether the application of exogenous spermidine (Spd) increases antioxidant activities and, therefore, elevates the heat tolerance of cucumber. Cucumber seedlings (cv. Jinchun No. 4) showing moderate heat tolerance were grown in climate chambers to investigate the effects of exogenous Spd (1 mm) foliar spray treatment on the activities and isozyme levels of antioxidative enzymes under both high-temperature stress 42/32 °C (day/night) and normal temperature 28/18 °C (day/night). On high-temperature stress, the activities of superoxide dismutase and ascorbate peroxidase were significantly reduced; the catalase activity was initially lower and then increased, whereas the peroxidase activity was initially higher and then decreased. The levels of these isozymes also changed differently. On treatment with exogenous Spd, the activities of these antioxidant enzymes were noticeably enhanced, and the isozyme zymogram expression had some changes. It was concluded that foliar spray with Spd effectively improved the total antioxidant ability of cucumber seedlings and, therefore, enhanced the tolerance of the plants to high-temperature stress.
Yan Yao, Yao Kong, Ping Zhang, Hua Zhang, Hong-di Huang and Guang-guang Li
The rapid expansion of Asian populations in the United States presents significant requirements for Asian vegetables. Flowering chinese cabbage (Brassica rapa L. ssp. chinensis var. utilis Tsen et Lee) is one of the most popular vegetables in China. The main factors restricting the progress in its breeding and genetic studies is the time required in generating desired pure line populations. Doubled haploid (DH) populations of flowering chinese cabbage have not been established because of technical difficulties. An appropriate combined protocol for a fast generation cycling system could advance up to seven generations, allowing the production of pure line seeds within 336–420 days among four cultivars and one hybrid of flowering chinese cabbage. The previous six generation cycles were accelerated using the embryo culture plus soil method which bypassed seed maturation through in vitro culture of immature embryos and promoted plant reproduction under stressed conditions, then the seventh generation cycle was accomplished until mature seeds were harvested using the soil method. During the culture of immature embryos, 12-day-old embryos could germinate and develop successfully on a Murashige and Skoog medium (MS) medium (Murashige and Skoog, 1962) containing 10% young coconut juice. This combined protocol bypasses the current obstacles in constructing DH populations of flowering chinese cabbage and is a possible alternative for producing pure lines. Its wider adoption could facilitate the breeding and biological studies of other Brassicaceae vegetables.
Yan-Chang Wang, Lei Zhang, Yu-Ping Man, Zuo-Zhou Li and Rui Qin
Big fruit size and nice red pigmentation combined with good flavor should be the major target for red-fleshed kiwifruit (Actinidia spp.) breeding programs. Genetic diversity and plant characteristics were evaluated on a set of kiwifruit accessions with predominantly red flesh to identify the superior individuals for further breeding or study of commercial application. The leading phenotypic characters varied widely among the accessions. Accession R reached average fruit weight ≈100 g, whereas it ranged from 43.15 to 84.71 g for the other accessions. Fruits of L and Q were flatter in shape than the others. The core volume accounted for fruit proportions ranging from 2.33% to 11.42%. ‘Chuhong’, ‘Honghua’, and K exhibited a round fruit apex, whereas most others showed a depressed apex. R, L, and Q had the highest a* values in the inner pericarp and also the most appealing visual coloration. Results revealed significantly higher soluble solid content (SSC), total sugar, and sugar/acid ratio in Q, R, and L. The 12 pairs of simple sequence repeat (SSR) markers were successfully used to characterize the genetic variability and confirm true-to-type identity for four accessions. However, the limited number of markers had no ability to discriminate among the other 11 accessions. Based on additional 28 SSRs, six of the indistinguishable accessions were confirmed to be genetically different, and three seemed to belong to the same clone vine. The results demonstrated that application of SSR data could improve the efficiency of identifying red-fleshed kiwifruit germplasm.
Zhi-Rong Li, Kang-Di Hu, Fen-Qin Zhang, Shi-Ping Li, Lan-Ying Hu, Yan-Hong Li, Song-Hua Wang and Hua Zhang
Broccoli (Brassica oleracea var. italica) is an important vegetable crop rich in vitamins and sulforaphane. However, the floral heads of broccoli experience rapid postharvest senescence. Here we found that hydrogen sulfide (H2S) treatment alleviated dark-promoted senescence in broccoli florets. H2S delayed the symptoms of senescence and maintained higher levels of chlorophyll and Rubisco and lower protease activity compared with water control. Gene expression analysis showed that H2S down-regulated the expression of chlorophyll degradation-related genes BoSGR, BoNYC, BoCLH1, BoPPH, and BoRCCR. Expression of lipoxygenase gene BoLOX1 and the genes involved in the ethylene synthesis pathway, BoACS2 and BoACS3, were also down-regulated by H2S. The reduced expression level in cysteine protease gene BoCP3 and aspartic protease gene BoLSC807 suggested the role of H2S in alleviating protein degradation during broccoli senescence. H2S up-regulated the expression of sulfur metabolism genes BoSR and BoOASTL, and the antioxidant gene BoCAT. These results show that H2S plays a vital role in alleviating broccoli senescence through a broad regulation on gene expression of reactive oxygen species (ROS) metabolism genes, ethylene synthesis genes, and protease genes.
Li-Qiang Tan, Xin-Yu Wang, Hui Li, Guan-Qun Liu, Yao Zou, Shen-Xiang Chen, Ping-Wu Li and Qian Tang
Landrace tea populations are important recourses for germplasm conservation and selection of elite tea clone cultivars. To understand their genetic diversity and use them effectively for breeding, two traditional landrace tea populations, Beichuan Taizicha (BCTZ) and Nanjiang Dayecha (NJDY), localized to northern Sichuan, were evaluated for morphological characters, simple sequence repeat (SSR)–based DNA markers and the contents of biochemical components. A wide range of morphological variation and a moderately high level of DNA polymorphism were observed from both BCTZ and NJDY. NJDY had on average, bigger leaves, larger flowers, higher total catechins (TCs), and greater gene diversity (GD) than BCTZ. Interestingly, samples from BCTZ had a wide range in the ratio of galloylated catechins to nongalloylated catechins (G/NG) (1.83–8.12, cv = 48.8%), whereas samples from NJDY were more variable in total amino acid (TAA) content (25.3–50.8 mg·g−1 dry weight) than those from BCTZ. We concluded that the two Camellia sinensis landrace populations are of great interest for both individual selection breeding and scientific studies.