Min-Li Liu, Ching-Hsiang Hsieh and Yun-Yang Chao
Renwei Huang, Daofeng Liu, Min Zhao, Zhineng Li, Mingyang Li and Shunzhao Sui
Lobularia maritima (L.) Desv. is an important ornamental plant. We investigated an efficient method to induce tetraploid plants of L. maritima (L.) Desv. by treating germinating seeds and apical growing points of seedlings with a range of concentrations of colchicine for different periods of time. Examination of the ploidy level by counting chromosome numbers at metaphase confirmed that the chromosome number of diploid plants was 2n = 2x = 24, whereas 2n = 4x = 48 was observed in tetraploid plants. The morphological characteristics of the diploid and colchicine-induced tetraploid plants were compared. Increases in the size of leaves, flowers, and stomata were observed in the tetraploid plants compared with the diploids. However, the stomatal density and plant height of the tetraploid plants were lower than for the diploid plants. This study presents the first report of autotetraploid plants of L. maritima (L.) Desv., and of the successful generation of tetraploid plants with improved ornamental traits by colchicine treatment.
Xiu Cai Fan, Hai Sheng Sun, Ying Zhang, Jian Fu Jiang, Min Li and Chong Huai Liu
In this study, simple sequence repeat (SSR) and sequence-related amplified polymorphism (SRAP) markers were used to analyze the genetic diversity of 48 wild Vitis davidii accessions. A total of 78 distinct alleles were amplified by 11 SSR primers, and the average allele number was 8.8. The average observed heterozygosity (Ho) and expected heterozygosity (He) values were 0.785 and 0.814, respectively. The effective allele numbers ranged from 3.92 to 9.61. The average polymorphism information content (PIC) was 0.798. Twelve of 169 SRAP primer combinations were selected for SRAP analysis. A total of 188 bands were produced, and the average was 15.7 bands per primer combination; the average percentage of polymorphic bands was 84.0%. The average PIC was 0.76. The results of the clustering analysis based on SSR markers showed that the 48 wild V. davidii accessions could be classified into five main clusters and had a genetic similarity coefficient level of 0.68. The dendrogram obtained from the SRAP data showed that 48 wild V. davidii accessions could be classified into five main clusters and had a genetic similarity coefficient of 0.72. SSR and SRAP markers differentiated all accessions studied including those with a similar pedigree. We speculated on the origin of Ciputao 0941♀, Ciputao 0940♂, and Fu’an-ci-01 using SSR markers and used both SSR and SRAP markers to resolve homonymy. The result will be valuable for further management and protection of V. davidii germplasm resources.
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.
Muhammad Irshad, Hafiz Muhammad Rizwan, Biswojit Debnath, Muhammad Anwar, Min Li, Shuang Liu, Bizhu He and Dongliang Qiu
The regeneration frequency of okra (Abelmoschus esculentus) is greatly influenced by its genetic makeup and recalcitrant nature. Phenolic secretion, in particular, is a major problem in okra tissue culture. This study describes a reproducible, rapid, and more efficient in vitro regeneration method using cotyledonary node explants of okra. Explants were incubated on Murashige and Skoog (MS) medium containing different concentrations and combinations of various plant growth regulators (PGRs) [benzyladenine (BA), thidiazuron (TDZ), and α-naphthylacetic acid (NAA)], and regeneration enhancers [silver nitrate (AgNO3) and Pluronic F-68]. Cut ends of cotyledonary node segments rapidly turned brown and cultures failed to establish. Antibrowning additives, such as activated charcoal (AC), ascorbic acid (AA), and AgNO3 at various concentrations in PGR-free MS basal medium were tested for their ability to control phenolic secretion from explants. Among these additives, 15 mg·L−1 AA was found to be optimal for controlling phenolic secretion, resulting in healthy explants and culture establishment. The highest number of shoots (a mean of 9.3 ± 0.9 shoots per cotyledonary node explant) was obtained on MS media containing 0.5 mg·L−1 NAA + 1 mg·L−1 TDZ + 0.1% Pluronic F-68. Individual shoots were elongated on MS medium + 1 mg·L−1 BA + 0.1 mg·L−1 gibberellic acid (GA3) (shoot length 5.3 ± 0.2 cm) and rooted on ½ MS medium + 1 mg·L−1 indole-3-butyric acid (IBA) and 200 mg·L−1 AC (5.3 ± 0.2 roots per shoot). Rooted plantlets were acclimatized in plastic pots inside a plant growth chamber at 25 ± 2 °C and 70% relative humidity, with an 80% survival rate. This optimized protocol can be used for producing transgenic plants of commercial okra cultivars through genetic transformation.
Ting Min, En-chao Liu, Jun Xie, Yang Yi, Li-mei Wang, You-wei Ai and Hong-xun Wang
Ethylene response factor (ERF) genes have been involved in responses to biotic and abiotic stress, including hypoxia and anaerobic stress. Vacuum packaging (a typical anaerobic stress) is an effective storage method used to delay browning of fresh-cut lotus root (Nelumbo nucifera). In model plants, ERF genes have been identified as responsive to hypoxia. Whether ERF is associated with browning of vacuum-packaged lotus root has not been studied. The effects of vacuum packaging on browning, phenolic content, the enzyme activity of phenylalanine ammonia lyase (PAL), polyphenol oxidase (PPO), and peroxidase (POD), and PPO, PAL, POD, and ERF genes expression in fresh-cut lotus root were studied. Downregulation of NnPAL1, NnPPOA, and NnPOD2/3 attributable to vacuum packaging coincided with increased related enzyme activities and the degree of browning of fresh-cut lotus root. The expression patterns of NnERF4/5 were consistent with the changes in NnPAL1, NnPPOA, and NnPOD2/3 gene expression. It has been proposed that NnERF4/5 could have be important regulators of fresh-cut lotus root browning, and that the relationships of NnERF4/5 and NnPAL1, NnPPOA, and NnPOD2/3 should to be studied further.