Zelkova sinica Schneid. is a popular landscape plant in China because of its wide adaptation, strong disease resistance, large crown, and beautiful fall color. Immature embryos from Z. sinica seeds were cultured on woody plant medium (WPM) supplemented with 4.5 μM 6-Benzylaminopurine (BA) and 5.4 μM α-naphthaleneacetic acid (NAA) to induce callus, and 60% of immature embryos formed callus. The cream-white, friable, nodular callus with proembryogenic structures was then cultured on WPM containing 5.4 μM NAA in combination with 9.0 or 11.2 μM BA to regenerate shoots; approximately five shoots per explant were induced on 70% callus. Shoots were rooted on WPM containing 0.5 μM indole-3-butyric acid (IBA), on which 62.3% shoots developed roots with an average of 4.2 roots per shoot at 4 weeks. The regenerated plantlets were acclimatized and transplanted into the field. This protocol could be used for mass production for field plantation, genetic improvement, and germplasm exchange of Z. sinica.
Xiaoling Jin, Xijun Hu, Youping Sun, Donglin Zhang and Ping He
Yu Zong, Ping Sun, Xiaoyan Yue, Qingfeng Niu and Yuanwen Teng
Pyrus betulaefolia is one of the most popular pear (Pyrus) rootstocks in China and other east Asian countries because of its good adaptability to versatile environments. However, the number of wild P. betulaefolia populations is decreasing because of habitat destruction and fragmentation. An urgent evaluation of P. betulaefolia genetic diversity and population structure is necessary to develop a conservation strategy for this important wild species. Thirteen simple sequence repeat loci were detected to infer the genetic composition of 18 P. betulaefolia populations in northern China. The average number of different alleles for each locus was 7.1. The number of effective alleles among loci ranged from 1.77 to 5.94. The overall mean values of expected and observed heterozygosity were 0.702 and 0.687, respectively. The Taihang Mountains, which run from northeast to southwest, acted as natural boundary in shaping the genetic diversity of P. betulaefolia in northern China. The distinct pattern, which was also observed in the distribution of chloroplast DNA (cpDNA) variation, appeared to be obscured by pollen-mediated gene flow in the distribution of nuclear microsatellite variation. Large populations with high allelic richness (e.g., populations BT, ZN, and QS) are considered suitable for in situ conservation because of the potential for adaptation to future environmental change. The smaller populations had mixed gene pools (e.g., populations GQ and XF) and should therefore also be considered for ex situ conservation. Preserving genetic diversity in seeds was proposed when field collections are fully characterized.
Chun-Qing Sun, Zhi-Hu Ma, Guo-Sheng Sun, Zhong-Liang Dai, Nian-jun Teng and Yue-Ping Pan
Reproductive barriers exist in some water lily crosses that result in low seed set and low breeding efficiency. We investigated pollen morphology, pollen viability, microspore development, pistil receptivity, and embryo and endosperm development in six water lily crosses using paraffin section as well as light and scanning electron microscopy (SEM) techniques. The results indicated that the percentage of pollen with normal morphology ranged from 8.9% to 55.2%. The pollen viabilities of ‘Fen Zhuang’, ‘Bai Lu’, and ‘Hong Ying’ were 33.9%, 3.3%, and 20.7%, respectively. Stigmatic pollen germination peaked at 12 h after pollination and varied from 0.3 to 65.7 grains per stigma among the crosses. The production of embryos with normal morphology ranged from 0% to 43.6% at 5 days after pollination, from 0% to 31.4% at 15 days after pollination, and from 0% to 19.7% by 20 days after pollination. The seed sets of the six crosses were from 0% to 10.9%. Our results suggest that the low seed set in some crosses is the result of low pollen viability, low pistil receptivity, and embryo abortion.
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.
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 220.127.116.11)] 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).