7(4) 2014 issue
July 2014 issue
Southern Cross Publishing Group©2014

Plant Omics | July 2014
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Proteomic analysis of the cold stress response in the leaves of birch (Betula platyphylla Suk)

Feng-Zhang WU1*, Bai-Chen WANG2 , Chuan-Ping YANG3

Modern Agriculture Research Academy of Dalian University, Dalian 116622, China

2Key Laboratory of Photobiology, the Chinese Academy of Sciences, Beijing 100093, China
3State Key Laboratory of Tree Genetics and Breeding (Northeast Forestry University), 26 Hexing Road, Harbin 150040, China

To investigate how actively growing birch (Betula platyphylla Suk) adapts to cold stress, two and a half-month-old birch plants were exposed to cold stress (4
°C) and compared to plant material maintained at the control temperature (23°C). This treatment did not affect the survival of the plants, but growth was almost arrested. Proteomic analysis, by bidimensional electrophoresis was performed, and a total of 15 protein spots were identified by mass spectrometry after 14 days of cold stress at 4°C. Proteins that were identified were involved in defense, photosynthesis, biosynthesis, carbon (C)-nitrogen (N) metabolism, and signal transduction. These proteins may be used for the establishment of a new network based on cooperation when plants are subjected to cold stress. Cold stress response proteins such as the b subunit of ATP synthase were mostly related to all aspects of chloroplast physiology, indicating that the cold resistance of birch was influenced, in part at least, by the chloroplast function. It was further found that the protein spots involved in defense responses, biosynthesis, C-N metabolism, and signal transduction were increased in intensity after cold stress, indicating that they played a key role in the cold hardiness mechanism of birch.

Pages 195-204 | Full Text PDF
Differential expression of antioxidant proteins in the drought-tolerant cyanobacterium Nostoc flagelliforme under desiccation

Wenyu Liang1, Lingxia Wang2, Jing Shi1, Xiaoting Lei1, Jia Yang1, Shijie Wu1, Wei Chen2, 3*

1School of Life Sciences, Ningxia University, Yinchuan, PR China
2Center for Molecular Cell and Systems Biology, School of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou, PR China
3Fujian Provincial Key Laboratory of Crop Molecular and Cell Biology, Fujian Agriculture and Forestry University, Fuzhou, PR China

Nostoc flagelliforme is a terrestrial nitrogen-fixing cyanobacterium that is distributed in arid or semi-arid steppes in Western and North-western of China. This species shows a strong ecological adaptability to xeric drought environments. 2DE coupled with MALDI-TOF-TOF/MS was used to analyze the differentially expressed antioxidant proteins in N. flagelliforme under desiccation. A total of five antioxidant proteins (eight protein spots) were successfully identified, including peroxiredoxin (Prx), Mn-containing catalase (Mn-CAT), iron superoxide dismutase (Fe-SOD), superoxide dismutase (SOD) and ferritin
. These proteins were all significantly down-regulated except for Prx that was up-regulated, when N. flagelliforme was subjected to 24-h desiccation. The colonies maintained very limited amounts of water (15%) and very low physiological activity to resist severe drought. These proteins may serve as biochemical markers for N. flagelliforme colonies under desiccation because of their significant changes in expression level.

Pages 205-212 | Full Text PDF | Supplementary Data
Genetic variation of hazel (Corylus heterophylla) populations at different altitudes in Xingtangsi forest park in Huoshan, Shanxi, China

XiaoYao Di, KeWu Liu, SaiQi Hou, PengLiang Ji, YiLing Wang *

College of Life Science, Shanxi Normal University, Linfen 041004, P.R. China

Corylus heterophylla, which grows along an altitude gradient ranging from 1200 m to 2400 m, is an important economic shrub species that helps maintain vegetation stability and ecological equilibrium in Xingtangsi forest park in Huoshan, Shanxi. In this study, eight C. heterophylla populations were sampled along an altitude gradient (each sampling interval was about 100m). By using a SRAP molecular marker, the genetic diversity and variation of C. heterophylla populations in Xingtangsi forest park were studied. The results showed that the C. heterophylla populations had high genetic diversity.

Pages 213-220 | Full Text PDF
Effects of cold acclimation on proteome expression patterns related to freezing tolerance in a Tibetan alpine plant Saussurea laniceps

Meng Yu1, Yuzhen Chen1*, Xiaozhong Lan2, Deshan Jin1, Junqi Zhang1, Cunfu Lu1*

1College of Biological Sciences and Biotechnology, National Engineering Laboratory for Tree Breeding, Key Laboratory of Genetics and Breeding in Forest Trees and Ornamental Plants, Ministry of Education, Beijing Forestry University, Beijing 100083, China

2Agricultural and Animal Husbandry College, Tibet University, Linzhi, Tibet 860000, China

Saussurea laniceps is a perennial herbal alpine plant possessing strong cold tolerance. To get a deeper insight into its cold tolerance mechanisms
, freezing tolerance and the proteomic profiles of cold-acclimated S. laniceps plantlets were analyzed. The survival rate of plantlets with height of 8-9 cm were recorded after exposure to chilling stress (2şC) and compared to plant material kept at a control temperature (23şC). The survival percent at -10°C increased from 0% to 40% during the 12 d of acclimation. Proteomic analyses, by two-dimensional gel electrophoresis (2-DE), performed during this stage revealed that 9 proteins were up-regulated, while 5 proteins were down-regulated. Among the proposed accumulating or appearing proteins, chlorophyll a-b binding protein 2 (LHCP-2), ribulose-1, 5-bisphosphate carboxylase/oxygenase activase (RCA), 33kDa manganese stabilizing chloroplast protein (33kDa MSP), and oxygen-evolving complex protein 1(OEC-1) were all related to photosynthesis, Maturase K is involved in gene expression regulation and galactinol synthase (GS) related to raffinose synthesis. Cold exposure induced a decrease in the candidate proteins including microtubule plus-end binding protein, ribosomal protein S13, O-acetylserine (thiol) lyase and photosystem I assembly protein ycf4. These results suggest changes in proteins associated with energy production, microtubule dynamic, raffinose synthesis and gene expression regulation process allow S. laniceps to enhance its freezing tolerance in the chilling environment.

Pages 221-228 | Full Text PDF
Proteomic analysis of the heat stress response in leaves of two contrasting chrysanthemum varieties

Yang Zhang1,2,3, Ming Sun1,2,3,*, Qixiang Zhang1,2,3

1College of Landscape Architecture, Beijing Forestry University, Beijing, 100083, China
2Beijing Key Laboratory of Ornamental Plants Germplasm Innovation and Molecular Breeding, Beijing, 100083, China
3National Engineering Research Center for Floriculture, Beijing, 100083, China 

High temperature stress causes damages to chrysanthemum by adversely affecting its growth and development. To identify differentially expressed proteins under heat stress, heat-tolerant variety (Chrysanthemum nankingense ) and heat-susceptible variety (Chrysanthemum ‘Wanfen’) were selected to contrast the full leaf proteome profiles with or without 6 h of heat stress (40°C), a parameter established in our previous study. Isobaric tags for relative and absolute quantitation (iTRAQ) was used in combination with tandem mass spectrometry and multidimensional liquid chromatography technology. Results of physiological tests showed that the variation of relative ion leakage and malondialdehyde were larger in Chrysanthemum ‘Wanfen’ than those in Chrysanthemum nankingense. In proteomics experiments, 250 proteins were identified, of which 43 were differentially expressed. Functional classification indicated that most of proteins were involved in photosynthesis, metabolic processes, oxidation-reduction processes and transport. Furthermore, 12 differentially expressed proteins were analyzed at the mRNA level under different durations (0 h, 3 h and 6 h) of heat treatment by qRT-PCR. The results showed that under heat stress the expression patterns of the rest 11 genes coding the altered proteins were correspondent to their respective protein levels with the exception of actin.

Pages 229-236 | Full Text PDF | Supplementary Data xls
Appropriate choice of antibiotics for plant regeneration and optimization of selective agents to be used in genetic transformation of chrysanthemum

Aung Htay Naing1, Kyung Il Park2, Sun Hyung Lim3, Chang Kil Kim1*

1Department of Horticultural Science, Kyungpook National University, Daegu 702-701, Korea
2Department of Horticulture & Life Science, Yeungnam University, Gyeongsan 712-749, Korea
3National Academy of Agricultural Science, Rural Development Administration, Suwon 441-707, Korea

This study was conducted to examine the effects of three antibiotics (carbenicillin, cefotaxime, and Clavamox) on in vitro plant regeneration from leaf segments of Chrysanthemum morifolium (Ramat.) ‘Vivid Scarlet’, and percentages of shoot induction and number of shoots per explant were recorded after 5 weeks of culture. Carbenicillin and Clavamox had less inhibitory effects on number of shoots per explant than cefotaxime, but superior plant growth (i.e. number of leaves, number of roots, plant height, and fresh weight) was observed in shoots treated with 125 mg
.L-1 Clavamox.

Pages 237-243 | Full Text PDF
Comparison and analysis of transcriptome profiles of haploid and diploid Populus by digital gene expression

Xuliang Mao1, Xuemei Xu2, Haizhen Zhang1, Jingli Yang1,3*, Zhanchao Wang1, Chenghao Li1,*

1State Key Laboratory of Forest Genetics and Tree Breeding, Northeast Forestry University, 26 Hexing Road, Harbin 150040, China
2Library of Northeast Forestry University, 26 Hexing Road, Harbin 150040, China

Haploid (HP) plants have been used for practical and basic research for many years. The development of haploid plants of Populus simonii Carr. × P. nigra L. was inhibited compared with the wild-type (diploid). In order to understand the molecular mechanisms and the genes expression characteristics, we performed gene expression profiling of the shoot tips and leaves of haploid and diploid plants using high-throughput tag-sequencing (Tag-seq) analysis, based on an Illumina Digital Gene Expression (DGE) platform. Approximately, 3.0-3.4 million total clean sequence tags with 0.09-0.1 million distinct clean tag sequences were obtained for each library. The clean tags were mapped to the reference sequences for annotation of expressed genes. About 53.93-56.64% of the distinct clean tags were mapped to the reference genes. Differentially expressed genes between haploid shoot tip library and diploid shoot tip library and differentially expressed genes between haploid leaf library and diploid leaf library were identified (4464 and 3856, respectively).

Pages 244-252 | Full Text PDF | Supplementary Data
Identification of TRAP and SRAP markers linked with yield components under drought stress in wheat (Triticum aestivum L.)

Khaled A. Moustafa1, M. Saleh2, A. A. Al-Doss1, A. A. Elshafei13, A. K. Salem1,4, F. H. Al- Qurainy2, M. N. Barakat1,5*

1Plant Production Department, College of Food and Agriculture Sciences, King Saud University, Riyadh, Saudi Arabia
2Botany and Microbiology Department, College of Pure Science, King Saud University, Riyadh, Saudi Arabia
3Genetics and Cytology Department National Research Centre, El-Dokki, Cairo, Egypt
Field Crops Research Department, National Research Centre, El-Dokki, Cairo, Egypt
5Biotechnology Laboratory, Crop Science Department, Faculty of Agriculture, University of Alexandria, Egypt

In order to identify TRAP and SRAP markers linked to yield components under drought stress in wheat (Triticum aestivum L.), a segregating F4 population from the cross between drought-sensitive (Yecora Rojo) and drought-tolerant (Pavon 76) genotypes was generated. The parents and 150 F4 families were evaluated phenotypically for drought tolerance using two irrigation treatments [2.5 and 7.5 m3(H2O) m-2(soil)]. The polymorphism among parental genotypes and F4 families were tested using 40 and 98 different TRAP and SRAP primer combinations, respectively. The results revealed that ten of fourteen TRAP and nine of nineteen SRAP markers are linked to the agronomic traits and can be introduced as new markers. The 14 TRAP markers were assigned to chromosomes 2A, 4A, 5A, 1B, 3B, 6B and 2D. The 19 SRAP markers were assigned to chromosomes 2A, 4A, 5A, 7A, 1B, 2B, 3B, 1D and 3D. Results also showed that all of the QTLs had a positive additive effect on agronomic traits, indicating contribution of alleles by the tolerant parent 'Pavon76'. QTLs for the six agronomic traits was associated with above mentioned markers and explained from 5 to 42 % of the phenotypic variation for all the agronomic traits. The genetic distance ranged from 10.1 to 38.1 cM. Therefore, the TRAP and SRAP markers linked to the QTL for the drought tolerance can be further used in breeding for drought tolerance in wheat.

Pages 253-259 | Full Text PDF | Supplementary Data
Genome-wide in silico identification and comparison of Growth Regulating Factor (GRF) genes in Cucurbitaceae family

Mehmet Cengiz Baloglu*

Kastamonu University, Faculty of Engineering and Architecture, Department of Genetics and Bioengineering, 37100, Kastamonu, Turkey

Growth-regulating factor (GRF) genes play a regulatory role for plant growth and development. The recently available cucumber, melon and watermelon genomes provide an opportunity to conduct a comprehensive overview of the GRF gene family. In the present study, identification and analysis of the GRF gene family was conducted using bioinformatics methods. Totally, 24 potential GRF genes were identified in cucumber, melon and watermelon. Cucumber and watermelon GRF gene members were physically mapped to their corresponding chromosomes. All GRF genes contain an intron whose number ranging from 2 to 3. Phylogenetic analysis categorized the cucurbit GRF proteins into 2 distinct classes. GRF proteins of cucurbits and Arabidopsis were clustered together in a joined tree and grouped into the same cluster with high bootstrap values.

Pages 260-270 | Full Text PDF
Trehalose-induced drought stress tolerance: A comparative study among different Brassica species

Md. Mahabub Alam1, Kamrun Nahar1,2, Mirza Hasanuzzaman3* and Masayuki Fujita1

1Laboratory of Plant Stress Responses, Department of Applied Biological Science, Faculty of Agriculture, Kagawa University, Miki-cho, Kita-gun, Kagawa 761-0795, Japan
2Department of Agricultural Botany, Faculty of Agriculture, Sher-e-Bangla Agricultural University, Sher-e-Bangla Nagar, Dhaka-1207, Bangladesh
3Department of Agronomy, Faculty of Agriculture, Sher-e-Bangla Agricultural University, Sher-e-Bangla Nagar Dhaka-1207, Bangladesh

Comparative responses of three Brassica species including B. napus, B. campestris and B. juncea under polyethylene glycol induced drought stress and the protective effects of exogenous Trehalose were investigated. Although drought reduced fresh, dry weight, chlorophyll (chl) contents; increased proline (Pro) content and oxidative stresses (lipoxygenase, LOX activity; malondealdehyde, MDA; H2O2 contents) along with altered antioxidant and glyoxalase systems in all Brassica species, B. juncea seems to be the most drought tolerant species showing the least oxidative damage due to enhancement of some non-enzymatic and enzymatic antioxidants. Combination of Tre and drought improved performance of all species, but responses were different. The results suggest that B. juncea is naturally drought tolerant species and moreover, its drought tolerance capability is further enhanced by exogenous Tre application.

Pages 271-283 | Full Text PDF
Effects of cadmium (Cd) on seedling growth traits and photosynthesis parameters in cotton
(Gossypium hirsutum L.)

Liantao Liu, Hongchun Sun, Jing Chen, Yongjiang Zhang, Dongxiao Li, Cundong Li*

Key Laboratory of Crop Growth Regulation of Hebei Province, Agricultural University of Hebei, Baoding 071001, China

Cadmium (Cd) is a widespread toxic heavy metal that usually causes deleterious effects in living organisms. In this study, the effects of Cd on seedling growth traits and photosyntheses parameters in cotton were investigated. The seedlings at three-leaf stage cultured in four nutrient solutions containing different Cd concentrations (0, 25, 50, and 100 µM) were subjected to assay the growth traits and the photosyntheses parameters. The growth traits analyzed included plant height, leaf area, biomasses of leaf, root, and stem, whereas the photosynthetic parameters measured included chlorophyll (Chl) content, gas exchange, and chlorophyll fluorescence. The results indicated that after 3 and 7 d treatments of Cd, the growth of cotton seedlings was significantly inhibited, showing that the plant height, biomass, and leaf area were all decreased. The Cd treatments also affected the photosyntheses parameters of the cotton seedlings. Under treatments of 50 and 100 µM concentrations of Cd, the seedlings exhibited significant decreases on Chl a and Chl b, photosynthetic rate (Pn), stomotal conductance (Gs), transpiration rate (E), maximal photochemical efficiency (Fv/Fm), quantum yield of electron transport (FPSII), photochemical quenching (qP), and electronic transport rate (ETR) (P=0.05). However, the Chl a/b and Ci were increased after the Cd treatments. The dramatic variations of chlorophyll fluorescence values under Cd treatments in comparison with the control (CK) suggested that the photosystem II (PS II) activity in cotton seedlings was much more sensitive to Cd toxicity. Taken together, our results confirm that Cd significantly inhibits the growth traits as well as the photosynthetic parameters in cotton seedlings. The non-stomata limitation, Chl reduction, and PSII system damage act as main limiting factors for affecting photosynthesis and cotton seedling growth under the Cd treatment.

Pages 284-290 | Full Text PDF