7(3) 2014 issue
May 2014 issue
Southern Cross Publishing Group©2014

Plant Omics | May 2014
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Proteomic evaluation of the response of soybean (Glycine max var Seoritae) leaves to UV-B

Sung-Eun Lee1,*, Sung Yung Yoo2,*, Do-Yeon Kim2, Tae Seok Ko2, Yong Sik Ok3, Tae-Wan Kim2,**

1School of Applied Biosciences, Kyungpook National University, Daegu 702-701, Korea
2Institute of Ecological Phytochemistry, Department of Plant Life & Environmental Science, Hankyong National University, Anseong 456-749, Korea
3Department of Biological Environment, Kangwon National University, Chuncheon 200-701, Korea

UV-B radiation plays an important role in photomorphogenesis; however, excessive UV-B radiation decreases photosynthesis and causes damage to cellular DNA. In the present study, two different light sources (UV-B and natural light) were applied to 18 day-old young soybean plants (Glycine max Merr. var Seoritae), after which the plants were harvested and their pigment contents, chlorophyll fluorescence, and proteomic changes investigated. The contents of carotenoids and anthocyanins increased significantly in response to excessive UV-B radiation. Additionally, several proteins such as ATP synthase, sedoheptulose-1,7-bisphosphatase, transketolase, peroxiredoxins and oxygen-evolving enhancer proteins were up-regulated in soybean leaves exposed to excessive UV-B. Alanine-2-oxoglutarate aminotransferase 1, aldolase, gamma-glutamyl hydrolase, and 28 kDa stem glycoprotein were down-regulated in soybean leaves exposed to excessive UV-B. Excessive UV-B light also led to a dramatic reduction in photosynthetic efficiency when compared to controls by causing irreversible damage to PSII determined by a fluorescence imaging system. With the treatment of natural illumination, the contents of carotenoids and anthocyanains were not changed in the leaves and the photosynthetic ability in the natural light was retained. These findings indicate that Seoritae soybean leaves might protect themselves from excessive UV-B radiation via up-regulation of antioxidative proteins and antioxidant pigments.

Pages 123-132 | Full Text PDF

In vivo
antibacterial activity against Rhizobium vitis and the induction of defense-related genes in grapevines (Vitis spp.) by hairy vetch and ryegrass extracts

Md. Tariqul Islam, SoonYoung Ahn, Md. Zaherul Islam, Seon Ae Kim and Hae Keun Yun*

Department of Horticulture and Life Science, Yeungnam University, Gyeongsan 712-749, Korea

Water based crude extracts from hairy vetch (Vicia villosa) and ryegrass (Lolium perenne) were investigated for their antimicrobial activity against Rhizobium vitis, the causal agent of crown gall of grapevine (Vitis spp.), in a greenhouse. A total of 100 ml of each extract (125, 250 and 500 g·L-1) prepared from fresh and pounded shoots and roots was applied to grapevine cuttings once every 10 days. The shoot and root extracts displayed remarkable in vivo antibacterial effects, as indicated by reductions in gall formation by R. vitis of 24.97% to 75.02% in ‘Kyoho’ (susceptible to crown gall) and 100% in ‘Campbell Early’(moderately resistant to crown gall) grapevines compared with the untreated control. The expression of several defense-related genes was investigated by quantitative real-time and semi-quantitative RT-PCR in two grapevine cultivars. Treatment with the extracts of hairy vetch and ryegrass led to differential induction of the accumulation of defense-related genes including
ß-1,3-glucanase (PR-2), phenylalanine ammonia-lyase (PAL), thaumatin-like protein (TLP), leucine reach-repeat (LRR), polygalacturonase inhibiting protein (PGIP), stilbene synthase (STS) and catalase (CAT). Expression analysis of defense related genes revealed noticeable differences between ‘Kyoho’ and ‘Campbell Early’ grapevines. Based on these results, extracts from tested cover crops can act as efficient biological inducers and therefore serve as an alternative strategy for grapevine crown gall protection.

Pages 133-141 | Full Text PDF

Proteomic analysis of Arabidopsis constitutive expresser of pathogenesis-related gene1 (Cpr30/cpr1-2) mutant

Xiaoqing Wang1,2, Mingyue Gou1,2, Huahu Bu3, Shengxue Zhang2, Guoying Wang2,*

1State Key Laboratory of Agrobiotechnology, China Agricultural University, Beijing 100193, China
2Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing 100081, China
3Research Centre of Modern Agriculture, Shanxi Academy of Agricultural Sciences, Taiyuan 030031, China

We have previously identified a gain-of-resistance mutant named constitutive expresser of pathogenesis-related gene1 (cpr30/cpr1-2). The mutant has dwarf morphology and gives rise to constitutive resistance to the bacterial pathogen Pseudomonas syringae in Arabidopsis. To elucidate the pattern of protein expression regulated by CPR1, we carried out a proteomic analysis of the cpr1-2 mutant compared with wild type (WT). Proteins of both 4-week-old WT and cpr1-2 plants were extracted by TCA-acetone-phenol and subjected to two-dimensional gel electrophoresis (2-DE) and matrix-assisted laser desorption/ionization-time-of-flight (MALDI-TOF) mass spectrometry. The results showed that the expression levels of some proteins were significantly affected in the cpr1-2 mutant, including pathogen defense-related proteins, redox-related proteins, energy metabolism proteins, signal transduction proteins, and proteins involved in protein folding and homeostasis. A majority of the identified proteins were related to reactive oxygen species (ROS) and some were central components of the unfolded protein response (UPR), suggesting that the mutation of CPR1 evoked oxidative stress and UPR signaling pathway activation, which may have contributed to the gain-of-resistance phenotype of cpr1-2. Some novel components of the differentially expressed proteins, such as GSR1, GB3, ClpC1, and ClpP5 that are rarely reported to be related to pathogen defense, were up-regulated in cpr1-2
, indicating new functions of these proteins or CPR1.

Pages 142-151 | Full Text PDF

Mapping of quantitative trait loci for aroma, amylose content and cooked grain elongation traits in rice

Acga Cheng1,2*, Ismanizan Ismail2,3, Mohamad Osman4 and Habibuddin Hashim5

1School of Biosciences, The University of Nottingham Malaysia Campus, 43500 Semenyih, Selangor Darul Ehsan, Malaysia
2 School of Biosciences and Biotechnology, The National University of Malaysia, 43600 Bangi, Selangor Darul Ehsan, Malaysia
3Institute of System Biology, The National University of Malaysia, 43600 Bangi, Selangor Darul Ehsan, Malaysia
4Faculty of Plantation and Agrotechnology, MARA University of Technology, 40450 Shah Alam, Selangor Darul Ehsan, Malaysia
5Malaysian Agricultural Research and Development Institute, 13200 Kepala Batas, Pulau Pinang, Malaysia

Global demand for high-quality rice has grown substantially in recent years and continues to trend upward due to taste preferences and a greater interest in healthy diets. Most rice quality traits are inherited in a complex way and may be affected by multiple genes and environmental factors. In this study, we conducted a simple sequence repeat (SSR)-based genetic analysis of quantitative trait loci (QTLs) affecting aroma, amylose content (AC) and cooked grain elongation (GE), which are the major quality characteristics in rice. The QTL analysis was performed using an F2 population of a cross between two Oryza sativa ssp. indica-type varieties, “Basmati 370” and “MR 84”, comprising of 90 plants. A total of 96 polymorphic markers were distributed over 12 rice chromosomes, each containing at least four markers. A total of nine QTLs (LOD>3.0) were detected for the three studied traits, including four for aroma, three for AC and two for GE. The highest number of QTLs was mapped to chromosome 8, with four QTLs for aroma. Two QTLs each for AC (qAC-3-1 and qAC-5-1) and GE (qGE-2-1 and qGE-2-2) are reported here for the first time. These QTL markers could be utilised as indirect selection tools in breeding programmes.

Pages 152-157 | Full Text PDF | Supplementary data
Review article

Research methods to study plant secondary metabolic pathways and their applications in the analysis of the biosynthetic pathway of stilbenes from Polygonum multiflorum -A review

Jiewen LI1, 2, Wei ZHAO1, Jian LI1, Wanxia XIA1, Lei LEI1, Shujin ZHAO1*

1Department of Pharmacy, General Hospital of Guangzhou Military Command, Guangzhou 510010, People’s Republic of China
2Guangzhou University of Chinese Medicine, Guangzhou 510006, People’s Republic of China

Plant secondary metabolites play significant roles in plant-environment interactions. Plant - derived secondary metabolites are compounds that can be widely used in industry, medicine, food sciences etc. In this review, we summarize the research methods applied to analyse biosynthetic pathways of plant secondary metabolites. Moreover we present an up to date overview of the studies that were performed on biosynthetic pathways of
stilbenes. In this manuscript we put emphasis on the biosynthetic pathway of 2,3,5,4'-tetrahydroxystilbene-2-O-ß-D-glucoside (TSG), an active component extracted from Polygonum multiflorum Thunb.

Pages 158-165 | Full Text PDF
Research Note

Molecular docking studies for discovery of plant-derived a-glucosidase inhibitors

Tae Kyung Hyun1, Seung Hee Eom1 and Ju-Sung Kim2,*

1Division of Applied Life Science, Plant Molecular Biology and Biotechnology Research Center, Gyeongsang National University, Jinju 660-701, Republic of Korea
College of Applied Life Sciences, the Research Institute for Subtropical Agriculture and Biotechnology, Jeju National University, Jeju 690-756, Republic of Korea

The identification of naturally occurring a-glucosidase inhibitors has been actively pursued with the aim of developing therapeutics for the treatment of type 2 diabetes mellitus. To identify a-glucosidase inhibitors, we screened 40 natural compounds, including flavonoids and phenolics, using the structure-based molecular docking approach. The rank of each compound was determined on the basis of the binding free energy of the lowest energy cluster. Results showed that all the tested compounds exhibited a binding energy ranging from -8.2 kcal/mol to -3.6 kcal/mol, indicating the variation in antidiabetic potential of tested compounds. The top-screened compounds were rutin, quercetin, and myricetin, which exhibited higher inhibitory activities (IC50 = 1.0 to 84.1 µg/mL) against a-glucosidase than acarbose (IC50 = 140.5 µg/mL), a reference inhibitor. We also demonstrated that a variety of inhibitory actions (competitive and noncompetitive inhibition) exist among these compounds. Our results suggest that rutin, quercetin, and myricetin can be used to further develop potent a-glucosidase inhibitors.

Pages 166-170 | Full Text PDF

Overexpression of the voltage-dependent anion channel 2 (VDAC2) gene induces drought resistance in Arabidopsis thaliana

Guoqin Wen*1,2, Yi Yang2, Liang Chai1,2,3, Zhibin Liu2, Jianmei Wang2

1College of Life Science, China West Normal University, Nanchong 637002, China
2Key Laboratory of Bio-Resources and Eco-Environment, Ministry of Education, College of Life Science, Sichuan University, Chengdu 610064, China
3Crop Research Institute, Sichuan Academic of Agricultural Sciences, Chengdu, 610066, China

Abiotic stresses, especially drought, trigger abscisic acid (ABA) signal conduction in plants. However, the molecular mechanisms are not well understood. We investigated the role of the voltage-dependent anion channel 2 gene (AtVDAC2) in drought resistance of Arabidopsis through ABA signalling. Some characteristics such as seed germination, root elongation, drought resistance, ABA related genes expressing, etc. were studied in plant transgenic lines. The AtVDAC2 overexpressing plants displayed an ABA-hypersensitive phenotype with higher seed germination and root elongation, while plants with AtVDAC2 knockdown (AtVDAC2-Dn) exhibited an ABA-insensitive phenotype. Additionally, AtVDAC2-OE plants showed enhanced resistance (about 40%) to drought than wide-type and AtVDAC2-Dn plants. In AtVDAC2 transgenic plants, some downstream targets in the ABA and/or drought-signalling pathways were altered at various levels, suggesting the involvement of AtVDAC2 in ABA-dependent drought resistance in Arabidopsis. Moreover, AtVDAC2-OE plants exhibited increased 9-cis-epoxycarotenoid dioxygenase 3 (NECD3) gene expression and generation of the reactive oxygen species hydrogen peroxide, suggesting that functional AtVDAC2 may be involved in ABA and H2O2 production process. The overexpression of the AtVDAC2 gene can confer drought resistance in Arabidopsis

Pages 171-177 | Full Text PDF

Deep sequencing-based transcriptome analysis of the oil-bearing plant Physic Nut (Jatropha curcas L.) under cold stress

Haibo Wang1,2, Zhurong Zou1, Shasha Wang1, Ming Gong1,*

1School of Life Sciences, Engineering Research Center of Sustainable Development and Utilization of Biomass Energy, Ministry of Education, Key Laboratory of Biomass Energy and Environmental Biotechnology of Yunnan Province, Yunnan Normal University, Kunming 650500, Yunnan, P. R. China
2College of Biological Resources and Environmental Science, Qujing Normal University, Qujing 655011, Yunnan, P. R. China

Nowadays Jatropha curcas L. has gained an increased attention in scientific and commercial fields as an important renewable bioenergy plant, aiming to prevent the possible energy crisis of fossil fuels. However, the studies on cold resistance of this biofuel shrub are still scarce, giving limited information for its genetic improvement and other biotechnological applications. In this work, the newly developed Illumina Hiseq™ 2000 RNA-seq, which is a deep high-throughput sequencing approach, preferentially was used for cold-resistance related transcriptome analysis of J. curcas. From the sequencing results, the total length of non-redundant sequences obtained was 4,960,092,780 nt, consisting of 106,749 contigs and 45,251 unigenes assembled by clean data. A total of 35,791 unigenes (79.09%) can be annotated to numerous databases (Nr, Swiss-Prot, GO, COG, KEGG) for functional classification. The 33,361 and 912 complete or partial CDSs are deduced by database alignment and ESTscan prediction, respectively. Among these unigenes, 27,293 can be categorized into 61 functional groups of GO, 11,887 of COG-annotated putative proteins were classified functionally into at least 25 molecular families, 18,787 were possibly involved in approximately 128 known metabolic or signaling pathways in KEGG. This study provided a comprehensive cold-resistance transcriptome analysis of J. curcas with remarkably more number of EST sequences than all previous relevant deposits in public databases. The results allowed us to decipher the key genes related to those coding for cold tolerance in this sequence library.

Pages 178-187 | Full Text PDF

Co-overexpression of AtCBL9, AtCIPK23 and AtAKT1 enhances K+ uptake of sugarcane under low-K+ stress

Qiwei Li1*, Lina Fan1, Qingwen Luo1, Huiyi He1, Jiachang Zhang2, Qiaoying Zeng1, Yu Li1, Wenling Zhou1, Zhenrui Huang1, Haihua Deng1, Yongwen Qi1*

1Guangdong Key Laboratory of Sugarcane Improvement and Biorefinery, Guangzhou Sugarcane Industry Research Institute, Guangzhou 510316, China
2State Key Laboratory of Plant Physiology and Biochemistry, College of Agronomy and Biotechnology, China Agricultural University, Beijing 100094, China

Low potassium (K+) availability is a major constraint to sugarcane growth and production. Previous studies have identified a calcium-signaling pathway that includes two calcineurin B-like calcium sensors (CBL1/CBL9) and a target interacting protein kinase (CIPK23) that activates the inward shaker-like potassium channel (AKT1), responsible for K+ uptake in plant roots. In this study, three components of the signaling pathway, AtCBL9, AtCIPK23 and AtAKT1 from Arabidopsis thaliana were co-overexpressed in sugarcane (Saccharum spp.) through particle bombardment. The results showed about 31% and 35% increase of K+ content in transgenic plants in low-K+ stress assay of tissue culture and hydroponic culture, respectively, compared to non-transgenic line. Under low K+ supply condition, transgenic lines have longer roots, higher plant height, heavier dry weight than non-transgenic line, indicating the transgenic lines grow better than non-transgenic lines. This study indicated that co-overexpression of AtCBL9, AtCIPK23 and AtAKT1 could significantly increase sugarcane K+ uptake ability and tolerance to low-K+ stress. These findings would have significant implications for improving stress tolerance of sugarcane in areas where K+ supply is low.

Pages 188-194 | Full Text PDF