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 Yao, Yao Kong, Ping Zhang, Hua Zhang, Hong-di Huang and Guang-guang Li
Jiang-Chong Wu, Jing Yang, Zhi-Jian Gu and Yan-Ping Zhang
By using a modified biotin-streptavidin capturing method, a total of 20 polymorphic microsatellite markers were developed from Moringa oleifera Lam. (Moringaceae), a useful multipurpose tree. Twenty-four domesticated individuals, with germplasms of India and Myanmar, were used to screen polymorphism of these 20 microsatellite markers. The number of alleles per locus ranged from two to six. The expected and observed heterozygosity varied from 0.3608 to 0.7606 and from 0.0000 to 0.8750, respectively. Seven loci were significantly deviated from Hardy-Weinberg equilibrium. The availability of these microsatellite primers would provide a powerful tool for aspects of detailed population genetic studies of M. oleifera.
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.
Chen Chen, Meng-Ke Zhang, Kang-Di Hu, Ke-Ke Sun, Yan-Hong Li, Lan-Ying Hu, Xiao-Yan Chen, Ying Yang, Feng Yang, Jun Tang, He-Ping Liu and Hua Zhang
Aspergillus niger is a common pathogenic fungus causing postharvest rot of fruit and vegetable, whereas the knowledge on virulence factors is very limited. Superoxide dismutase [SOD (EC 184.108.40.206)] is an important metal enzyme in fungal defense against oxidative damage. Thus, we try to study whether Cu/Zn-SOD is a virulence factor in A. niger. Cu/Zn-SOD encoding gene sodC was deleted in A. niger [MA70.15 (wild type)] by homologous recombination. The deletion of sodC led to decreased SOD activity in A. niger, suggesting that sodC did contribute to full enzyme activity. ΔsodC strain showed normal mycelia growth and sporulation compared with wild type. However, sodC deletion markedly increased the cell’s sensitivity to intracellular superoxide anion generator menadione. Besides, spore germination under menadione and H2O2 stresses were significantly retarded in ΔsodC mutant compared with wild type. Further results showed that sodC deletion induced higher superoxide anion production and higher content of H2O2 and malondialdehyde (MDA) compared with wild type, supporting the role of SOD in metabolism of reactive oxygen species (ROS). Furthermore, ΔsodC mutant had a reduced virulence on chinese white pear (Pyrus bretschneideri) as lesion development by ΔsodC was significantly less than wild type. The determination of superoxide anion, H2O2, and MDA in A. niger-infected pear showed that chinese white pear infected with ΔsodC accumulated less superoxide anion, H2O2, and MDA compared with that of wild type A. niger, implying that ΔsodC induced an attenuated response in chinese white pear during fruit–pathogen interaction. Our results indicate that sodC gene contributes to the full virulence of A. niger during infection on fruit. Aspergillus niger is one of the most common species found in fungal communities. It is an important fermentation industrial strain and is also known to cause the most severe symptoms in fruit during long-term storage (Pel et al., 2007). Meanwhile, plants activate their signaling pathways to trigger defense responses to limit pathogen expansion. One of the earliest host responses after pathogen attack is oxidative burst, during which large quantities of ROS are generated by different host enzyme systems, such as glucose oxidase (Govrin and Levine, 2000). ROS such as singlet oxygen, superoxide anion, hydroxyl (OH−), and H2O2 are released to hinder the advance of pathogens (Gara et al., 2003). ROS can react with and damage cellular molecules, such as DNA, protein, and lipids, which will limit fungal propagation in the host plant (Apel and Hirt, 2004).
Shuai-Ping Gao, Kang-Di Hu, Lan-Ying Hu, Yan-Hong Li, Yi Han, Hui-Li Wang, Kai Lv, Yong-Sheng Liu and Hua Zhang
Hydrogen sulfide (H2S) was recently recognized as an endogenous gaseous molecule involved in seed germination, root organogenesis, abiotic stress tolerance, guard cell movement, and delay of senescence in plants. In the present study, we show that H2S participates in the regulation of postharvest ripening and senescence in fresh-cut kiwifruit, Actinidia deliciosa. Fumigation of fresh-cut kiwifruit with the H2S donor sodium hydrosulfide (NaHS) solution prolonged kiwifruit storage time and alleviated senescence and tissue softening in a dose-dependent manner at an optimal concentration of 1.0 mmol·L−1 NaHS. H2S treatment maintained higher levels of reducing sugars, soluble proteins, free amino acids, ascorbate, and chlorophyll and lowered carotenoid levels. H2S treatment also significantly decreased the contents of malondialdehyde (MDA), hydrogen peroxide (H2O2) and superoxide anion (•O2 −) during fruit storage compared with water controls. Furthermore, the activities of guaiacol peroxidase (POD), catalase (CAT), ascorbate peroxidase (APX), and glutathione reductase (GR) were increased by H2S treatment, whereas the activity of lipoxygenase (LOX) was decreased compared with untreated controls. Taken together, these results suggest that H2S is involved in prolonging postharvest shelf life and plays an antioxidative role in fresh-cut kiwifruit.