Articles

7(5) 2014 issue
 
September 2014 issue
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Southern Cross Publishing Group©2014
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Plant Omics | September 2014
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Genome-wide analysis of the heat shock transcription factor family in Triticum urartu and Aegilops tauschii

Wenlong Yang, Juan Li, Dongcheng Liu, Jiazhu Sun, Lixiong He, Aimin Zhang*

Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, 100101, Beijing, China
College of Life Sciences, Hunan Agricultural University, 410128, Changsha, China

Abstract
Heat shock proteins (Hsps) are believed to play essential roles in developmental processes and in responses to heat stress. Heat shock transcription factors (Hsfs) are important Hsp regulators, but their functions are poorly understood, especially in wheat. In this study, a comprehensive bioinformatics analysis was conducted in wheat A and D genome donors, Triticum urartu and Aegilops tauschii, the genomic sequences of which were published recently. The results showed that 13 Hsf proteins were identified in both T. urartu and Ae. tauschii, and they could be classified into three groups according to structure; seven Hsfs belonged to group A, two to group B, and three to group C. Expression analyses of these Hsf genes in different tissues of T. urartu and in the response to heat stress were conducted using quantitative RT-PCR. Several Hsf genes in group A (Tuhsf03, Tuhsf05, Tuhsf06, Tuhsf10) had 19-292-fold increases in transcript levels versus the control in different tissues of T. urartu and could be induced by heat stress, while the transcripts of group B and group C Hsf genes could hardly be detected. These results provide important information for cloning, expression, and functional studies of Hsfs in wheat.

Pages 291-297 | Full Text PDF

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Isolation and expression of antimicrobial Camel lactoferrin (cLf) gene in tobacco

Seyyed Mohsen Sohrabi, Ali Niazi*, Mahmood chahardooli, Farzaneh Aram

Institute of Biotechnology, Shiraz University, Shiraz, Iran

Abstract
In this study, cDNA encoding the Camelus dromedarius lactoferrin (cLf) was isolated from mammary gland by reverse transcription polymerase chain reaction and cloned to the pART27 expression vector. Its predicted amino acid sequence was analyzed and compared with lactoferrin of other mammalian species. Secondary and tertiary structures were predicted and modeled by online tools. Then cLf cDNA was expressed in Nicotiana tobaccum cv. Xanthi using Agrobacterium mediate transformation. Analysis of transgenic plants by PCR, RT-PCR and Real-time PCR showed the recombinant camel lactoferrin gene was expressed in transgenic plants. Tests on protein extract from transgenic tobacco leaves demonstrated antimicrobial activity against microbial strains. Evaluations for inhibition efficiency against Staphylococcus aureus PTCC1112 (ATCC 6538), Escherichia coli PTCC 1330 (ATCC 8739), Bacillus subtilis PTCC1023 (ATCC 6633) and Candida albicans PTCC5027 (ATCC 10231) were 48.8%, 52.7%, 34.6% and 41.6% respectively. However, tests on protein extract from non-transformed plants did not display any sign of antimicrobial activity. Lactoferrin is a valuable protein that takes part in a large number of important physiological processes. Lf expression in plants provides a suitable system for large-scale production of this protein. Furthermore, it has been demonstrated that Lf expression in plants confers resistance to plant diseases, so this antimicrobial protein could also be used to improve crop quality.


Pages 298-307 | Full Text PDF
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MicroRNA*s in apple (Malus domestica): biological implications obtained from high-throughput sequencing data

Chaogang Shao, Xiaoxia Ma, Guofu Lu, Yijun Meng*

College of Life Sciences, Huzhou Teachers College, Huzhou 313000, P. R. China
College of Life and Environmental Sciences, Hangzhou Normal University, Hangzhou 310036, P. R. China

Abstract
MicroRNAs (miRNAs) play important roles in diverse biological processes in plants through the regulation of gene expression. Recently, the plant miRNA* species have been uncovered to possess potential biological functions through target cleavages, which is similar to the miRNAs. By utilizing the publicly available small RNA high-throughput sequencing data, we performed a comprehensive search for the miRNA* sequences on the currently annotated miRNA precursors of apple. This search was based on the secondary structures of miRNA precursors and the accumulation levels of the miRNA* candidates. As a result, 188 miRNA*s were identified on 170 pre-miRNAs in apple. The organ-specific accumulation patterns of the identified miRNA*s were investigated. The result shows that some of the miRNA*s are highly abundant in the apple leaves, roots, flowers or fruits, indicating the potential organ-specific functions of these miRNA*s. Several pieces of literature-based hints were obtained to link the organ-specific accumulation patterns of the miRNA*s to their potential biological roles in organ development. To gain deeper insights into the biological roles of the miRNA*s in apple, degradome sequencing data-based target identification was performed. The result shows that certain miRNA*s possess great potential of performing target cleavage-based regulation of gene expression. Taken together, our study could advance the current understanding on the regulatory activities of the miRNA*s in the non-model plant apple tree.

Pages 308-321 | Full Text PDF

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Effect of explant types and plant growth regulators on direct regeneration in medicinal plant Pogostemon cablin

Hua Jin, Zhi-Cheng Deng, Hong He
*

School of Chinese Materia Medica, Guangzhou University of Chinese Medicine, Guangzhou 51006, China

Abstract
The present work was conducted to develop a high frequency and rapid in vitro regeneration protocol for Pogostemon cablin (Blanco) Benth. In order to improve the regeneration efficiency of this crop, the effects of explant types and plant growth regulators (PGRs) on in vitro shoot proliferation, and also the subsequent rooting of shoots were examined. Of the explant types, nodal stem with a single node (the second or third node of in vitro plantlets) was the most responsive explant, with 100% explants producing 129.7-138.1 shoots on the optimal medium, and also leaf and petiole possessed a high regenerative capacity. Of the tested cytokinins, 6-Benzyladenine (BA) was most effective on shoot regeneration. BA at 0.1-0.2 mg·L-1, high number and length of shoots per explant were achieved. Combinations of BA and 1-napthaleneacetic acid (NAA) resulted in slower shoot development and growth as compared to BA alone. For regenerated shoots rooting, half-strength MS medium supplemented with 0.2 mg·L-1 indole-3-butyric acid (IBA) was most effective with maximum number (49.3 roots per plantlet) and length (1.45 cm per root) of roots respectively. Regenerated plantlets were successfully transferred into pots with soil and over 90% of them grew into healthy mature plants. The current study provided information toward commercial in vitro propagation of Pogostemon cablin.

Pages 322-327 | Full Text PDF

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Transcriptome analysis of differential expression genes from petals and lips of Phalaenopsis amabilis to identify genes associated with floral development

Yan-Xiao Li, Xiao-Ming Song, Zhen Wang, Shan-Wu Lv, Chang-Wei Zhang*

State Key Laboratory of Crop Genetics and Germplasm Enhancement/Key Laboratory of Biology and Germplasm Enhancement of Horticultural Crops in East China, Ministry of Agriculture, Nanjing Agricultural University, Nanjing 210095, China

Abstract
Phalaenopsis amabilis
belongs to the genus Phalaenopsis, which is member of Orchidaceae, one of the largest families of flowering plants. Orchidaceae includes species with greatly diversified and specialized floral morphology. But few genomic resources are available for these non-model plants. In order to understand the genetic mechanisms underlying floral patterning, we investigated differential expression genes from petals and lips of this species. Two libraries were prepared from petals and lips and then sequenced using short reads sequencing technology (Illumina) to identify the differential expression genes. The total reads were 8,889,080 and 16,224,038 for these two libraries, respectively. The open reading frame of unigenes was predicted using Getorf software. The MISA software was used to identify SSR markers for the unigenes that were larger than 1 Kb. Sequencing data between samples compared with the Unigene database, using soapsnp build consensus sequence, and then analyzed between samples to get homozygous SNP loci. By comparing the transcripts from petals and lips, we finally obtained 2,389 differentially expressed genes. These genes were significantly enriched in 101 KEGG pathways and 55 GO terms.  The transcriptome analysis provided a comprehensive understanding of the complexity of floral development and organ identity. The results let us know more details about the relationship between MADS gene family and floral morphology. This information broadens our understanding of the mechanisms of floral patterning and contributes to molecular and genetic research by enriching the Phalaenopsis database. 

Pages 328-336 | Full Text PDF | Supplementary Data PDF | Supplementary RAR
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Inhibition of plant pathogens in vitro and in vivo with essential oil and organic extracts of Torreya grandis ‘Merrilli’ aril

Xiaohong Sun, Nitin Mantri, Jian Ge, Yijun Du, Guofu Wang, Jiayin Lu, Wu Jiang, Hongfei Lu*

Shaoxing University Yuanpei College, Shaoxing, Zhejiang, 312000, China
School of Applied Sciences, Health Innovations Research Institute, RMIT University, Melbourne, 3000, Australia
College of Life Sciences, China Jiliang University, Hangzhou, 310018, China
Division of General Studies, University of Illinois at Urbana-Champaign, Champaign, IL, 61820, United States
College of Life science, Zhejiang Sci-Tech University, Hangzhou, 310018, China
College of Chemistry and Life Science, Zhejiang Normal University, Jinhua, 321004, China

Abstract
The seed aril of Torreya grandis ‘Merrilli’, a native conifer from China, has recently become popular for its food and pharmaceutical properties. The aims of this study were to comprehensively evaluate the efficacy of essential oil and other aril extracts as antifungal agents. The chemical components of the oil were analyzed by GC-MS. The results reveal the existence of more than 57 compounds in the volatile fraction, of which the main compounds were limonene (16.789%) and ί-pinene (8.553%). The aril oil and organic extracts showed remarkable antifungal property against five commercially important plant pathogenic fungi (Fusarium oxysporum f. sp. cucumerinum, Pyricularia oryzae, Bipolaris maydis, Pseudoperonospora cubensis, Alternaria solani). The EC50 ranged from 0.608-1.249 mg mL-1 and 0.241-1.350 mg mL-1, respectively. The oil had a strong detrimental effect on spore germination of all tested plant pathogens in vitro. Moreover, aril essential oil significantly reduced the damage areas produced by Pseudoperonospora cubensis and Alternaria solani on cucumber leaves and tomato fruits, respectively. The relative control efficiency (92.25%) was higher for essential oil treated cucumber leaves than that of Carbendazim treated leaves (84.27%). This is the first study that demonstrates T. grandis ‘Merrilli’ aril oil and its organic extracts possess a wide range of antifungal activity. They therefore have the potential of becoming an alternative to synthetic fungicides for controlling economically important plant fungal diseases.

Pages 337-344 | Full Text PDF

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Development of 19 transferable Cucurbita pepo EST-SSR markers for the study of population structure and genetic diversity in pumpkin (Cucurbita Moschata)

Weihua Mao, Yingjun Xu, Shengchun Xu, Lingwei Ye, Hui Wang, Xu Gao, Xiaorong Mo* and Yaming Gong*

Center of Analysis and Measurement, Zhejiang University, Hangzhou 310058, China
Cixi Agricultural Technology and Popularization Center, Cixi 315300, China
Institute of Vegetables, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China
State Key Laboratory of Plant Physiology and Biochemistry, College of Life Science, Zhejiang University, Hangzhou 310058, China

Abstract

Cucurbita moschata
is an economically important species worldwide, but little is known about its genic and genomic information. Few molecular markers have been developed for C. moschata so far, which is not sufficient for breeding programs. The aim of this study was to examine the feasibility of development EST-SSR markers from Cucurbita pepo to C. moschata and to evaluate their potential for genetic analysis in C. moschata cultivars. Eighty-two C. pepo EST-SSRs were tested against 29 C. moschata cultivars from different geographical origin in China. As a result, a high proportion (84.14%) of markers was transferable to C. moschata. From them, nineteen markers exhibited polymorphism. A total of 53 alleles were identified with an average of 2.789 alleles per locus in all the tested samples. The polymorphic information content (PIC), the observed heterozygosity (HO), the expected heterozygosity (HE), and the Shannon’s information index (I) were estimated to average 0.393, 0.125, 0.478, and 0.760, respectively. Population structure, principal coordinates and phylogenetic analysis all revealed that these 29 C. moschata cultivars tended to group into two subpopulations, which was related to fruit shape rather than geographic origin. Moreover, every subpopulation possessed its own population-specific alleles
. This is the first report describing the development of transferable EST-SSR markers in C. moschata and their application <http://dict.baidu.com/s?wd=application> to genetic analysis, which will offer an approach for future marker development and marker-assisted breeding in C. moschata.

Pages 345-352 | Full Text PDF

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Isolation and characterization of a phenylalanine ammonia-lyase gene (PAL) promoter from Ginkgo biloba and its regulation of gene expression in transgenic tobacco plants

Wei-Wei Zhang, Jin-Bao Li, Feng Xu*, Yin Tang, Shui-Yuan Cheng*, Fu-Liang Cao

Forest resources and Environment Institute, Nanjing Forestry University, Nanjing 210037, China
College of Horticulture and Gardening, Yangtze University, Jingzhou 434025, China
Huanggang Normal University, Hubei Key Laboratory of Economic Forest Germplasm Improvement and Resources Comprehensive Utilization, Huanggang 438000, China

Abstract
The Ginkgo biloba produces flavonoids and ginkgolides of high interest due to their medical values. Phenylalanine ammonia-lyase (PAL) is a core enzyme in the flavonoid biosynthesis pathway. In this study, we constructed genomic libraries with the DNA from leaves of 18-year-old grafts G. biloba. Using nested PCR method, a 1,627 bp 5’ flanking region, named GbPALp (GenBank: GU968736) of a PAL gene (GbPAL) was isolated from genomic libraries, and the analysis of the promoter sequence by the PLACE database has revealed the existence of several putative cis-elements. To assess the organ-specificity and developmental characteristics of PAL gene expression in G. biloba, the GbPALp-driven GUS expression in transgenic tobacco was studied. Histological analysis of the transgenic young tobacco plants showed that the cloned PAL promoter displayed a tissue-specific GUS staining restricted to root hair region, the cortex of root, the vascular bundle cells, the cortex and the primary xylem of stems, and vascular region of leaves. In transgenic mature plant, GUS was expressed in the spire apical meristem of stems but not in leaves and root. The GUS activity in transgenic young tobacco leaves was also observed to be induced by a variety of stresses, including UV-B, abscisic acid, methyl jasmonate and salicylic acid, respectively. The results indicated that GbPALp had multiple functions in the expression under the various developmental stages and stress conditions in the transgenic tobacco.

Pages 353-360 | Full Text PDF

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Physio-morphological responses of sweet potato [Ipomoea batatas (L.) Lam.] genotypes to water-deficit stress

Suravoot Yooyongwech, Thapanee Samphumphuang, Cattarin Theerawitaya, Suriyan Cha-um*

Department of Agricultural Science, Mahidol University, Kanchanaburi Campus, Kanchanaburi 71150, Thailand
National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA), 113 Thailand Science Park, Pahonyothin Road, Khlong Nueng, Khlong Luang, Pathum Thani 12120 Thailand

Abstract
Roots of sweet potato [Ipomoea batatas (L.) Lam.; Convolvulaceae] are rich source of carbohydrates, vitamins and other nutrients; however, root storage and productivity is very sensitive to water deficit stress. We, therefore, investigated the light harvesting complexes (photosynthetic pigments) and activities (chlorophyll fluorescence), and photosynthetic abilities in three genotypes of sweet potato in response to decreased soil water content (SWC). Single vine cutting was propagated and then water withheld in different soil water contents. Osmotic potential, free proline, chlorophyll pigments, chlorophyll fluorescence, net photosynthetic rate and growth characters were measured. Free proline in the leaf tissues was enriched depending on a degree of water deficit, genotypes and their interaction. Physio-morphological characteristics of water-deficit stressed plants in each genotype of sweet potato were significantly inhibited. Osmotic potential in leaf tissues of water-deficit stressed plants of Tainung 57 sharply declined (-0.044x) when compared to PROC 65-3 (-0.027x) and Japanese Yellow (-0.025x). Chlorophyll a (Chla), chlorophyll b (Chlb), total chlorophyll (TC), photon yield of PSII (
FPSII), stomatal conductance (gs), transpiration rate (E), vine length and number of leaves in cv. PROC 65-3 grown under water deficit were maintained better than those in cvs. Japanese Yellow and Tainung 57. A positive relationship between photosynthetic pigments and photosynthetic abilities was observed (R2 > 0.9) and it correlated directly with net photosynthetic rate (Pn). Free proline enrichment may play a key role as osmotic adjustment in sweet potato cv. PROC 65-3, grown under water deficit stress. Photosynthetic pigments, chlorophyll fluorescence activities, net photosynthetic rate and transpiration rate in cv. PROC 65-3 under water deficit condition were retained better than those in cvs. Japanese Yellow and Tainung 57, resulting in maintain growth performance.

Pages 361-368 | Full Text PDF

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Genetic analysis of extra glume and molecular mapping of eg3 in rice (Oryza sativa L.) using SSR markers

Xijuan Zhang*, Dan Liu*, Shukun Jiang, Jiayu Wang

Cultivation and Farming Research Institute, Heilongjiang Academy of Agricultural Sciences Postdoctoral research station, Heilongjiang Academy of Agricultural Sciences, 150086 Harbin, China
Key Laboratory of Crop Physiology, Ecology, Genetics and Breeding, Ministry of Agriculture, Shenyang Agricultural University, 110866 Shenyang, China

Abstract
The great advances of flower development have been achieved in the past decade by genetic and molecular analysis of floral homeotic mutants in Arabidopsis thaliana and Antirrhinum majus. But the developmental process of inflorescence and spikelet is still little known in rice, an important crop plant and model monocot plant. In this study, we isolated and characterized a novel extra glume3 (eg3) mutant, which produced extra glume-like structure organ on the lemma side of rice spikelet and developed between empty glumes and lemma. To analyze the inheritance of the eg3, we generated two segregating F2 populations by crossing eg3 mutant with normal cultivar Longdao5 and Qishanzhan. Our results confirmed that a single recessive nuclear gene controls the extra glume trait. Simple sequence repeat (SSR) and bulked segregant analyses of the F2 population revealed that the eg3 gene is located on chromosome 4. Using bulked-extreme and recessive-class approach, the eg3 was mapped between SSR marker RM471 and RM16842. According to the rice annotation project database, the target region is about 64.1 kb from 18,996,727 bp to 19,060,851 bp.

Pages 369-372 | Full Text PDF

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Characterization of terpene synthase gene family in Artemisia annua L.

Weizhang Jia, Ying Wang*, Xuesong Yu, Ling Zhang, Shunhui Liu, Zebo Huang*

School of Bioscience and Biopharmaceutics, Guangdong Pharmaceutical University, Guangzhou 510006, China
Guangdong Provincial Key Laboratory of Biotechnology Candidate Drug Research, Guangzhou 510006, China

College of Horticulture, South China Agricultural University, Guangzhou 510640, China

Abstract
Terpene synthases (TPSs) are responsible for the synthesis of the various terpene molecules which play important roles in the accumulation of secondary metabolites. However, systematic analysis of Artemisia annua TPS family has not been carried out, and little is known about the differences of transcript levels for TPS genes in A. annua (AaTPSs) and the regulation thereof. Though plant TPSs are generally divided into seven clades, a family of TPS in A. annua was characterized with lineages having a majority of TPS in TPS-a and TPS-b clades. Semi-quantitative RT-PCR demonstrates that 17 AaTPS genes are functional or potentially functional and express in at least some tissues or organs of the plant. In trichomes, the transcripts of majority of AaTPSs and artemisinin synthesis-related genes are express at a higher level than in leaves containing trichomes, except that AaBFS, AaCPS and AaSTS1 appear to be lower in trichomes but higher in leaves. AaECS is mainly express in trichomes, but although the highest expression level was detected in old leaf trichomes. The expression of AaSQS in leaves was slightly higher than that of trichomes, indicating that this gene is also express mainly in other parts of leaves instead of trichomes. Characteristics of sequence motifs, phylogenetic relationships and transcript levels of AaTPSs provide a foundation for future functional analysis. The present study contributes more data on the tissue-specific expression of AaTPSs in A. annua, and provides clues to dissect the concerted regulation for AaTPSs expression and the specific competitor for farnesyl diphosphate (FDP) in A. annua trichomes.

Pages 373-381 | Full Text PDF | Supplementary Data PDF

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Response of soybean plants to gamma radiation: Biochemical analyses and expression patterns of trichome development

Φzge Ηelik*, Ηimen Atak, Zekiye Suludere

Istanbul Kόltόr University, Faculty of Science and Letters, Department of Molecular Biology and Genetics, 34156, Atakφy, Istanbul, Turkey
Gazi University, Faculty of Science, Teknikokullar 06500, Ankara, Turkey


Abstract
This is the first report on soybean with the aim to show the effects of gamma radiation on trichome metabolim. Soybean seeds were subjected to 300 Gy gamma radiation at a dose rate of 10 Gy/min using a Cs-137 gamma source. The photosynthetic pigment, total protein content and ascorbate peroxidase activity were studied. The results shoed that the chlorophyll a content was decreased by 80% on day 14 and by 77% on day 21 of irradiation. The chlorophyll b content was reduced by 58.6% and 62.06% on day 14 and 21 after irradiation, respectively. The total carotenoid concentration was reduced by 81.14% on the 14th day after irradiation and by 91% on the 21st day of irradiation, compared to control. The total protein concentration was found to have decreased significantly at 14 and 21 day after treatment. High level of ascorbate peroxidase (APX) activity was recorded in the leaves developed from irradited soybean seeds, compared to the non-irradiated group. The trichome densities were 6.76 fold increased at 21 day of irradiation, while the stomatal densities were decreased, compared to control. We also performed A qRT-PCR analysis to detect the transcription levels of the soybean trichome developmental genes. The GL2 and CPC genes were up-regulated (P=0.05). The results of this study pointed out that the CPC transcription factor has to be study further studied to provide an insight on its exact role in regulation of trichome development in soybean under radiation stress.


Pages 382-391 | Full Text PDF
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Accumulation and tolerance characteristics of chromium in nine jute varieties (Corchorus spp. and Hibiscus spp.)

Muhammad Kamrul Islam, Iftekhar Alam, Mst Salma Khanam, Si Young Lee, Tatoba R Waghmode, Moo Ryong Huh*

Department of Horticulture, College of Agriculture and Life Science, Gyeongsang National University, Jinju 660-701, Korea
Institute of Agriculture and Life Science (IALS), Gyeongsang National University, Jinju 660-701, Korea
National Institute of Biotechnology, Ganakbari, Savar, Dhaka 1349, Bangladesh
Department of Agronomy, Bangladesh Agricultural University, Mymensingh 2302, Bangladesh


Abstract
This study investigates the effect of chromium (Cr) on seed germination, growth characteristics and the Cr accumulation in nine jute varieties. Two separate experiments viz, germination and vegetative growth were analyzed. Plants were exposed to 0-500 ppm Cr for 30 days and were evaluated for growth, gas exchange, photosynthetic activity, tolerance index and Cr accumulation. Excess chromium negatively affects on germination parameters such as the germination frequency (GF), germination index (GI) and vigor index (VI). There was a clear difference in germination properties across the varieties particularly in O-795 and O-9897. All varieties were moderately tolerant to Cr toxicity, with O-795, CVE-3, and BJC-7370, showed small inhibitions in plant growth and photosynthetic characteristics than the others. Furthermore, O-795, CVE-3 and BJC-7370 exhibited higher Cr concentration in the root, stem, leaf, higher bio-concentration factor as well as higher biomass than other jute varieties. Therefore, O-795, CVE-3 and BJC-7370 jute varieties can be used as excellent annual crop candidates for phytoremediation of Cr polluted soils.

Pages 392-402 | Full Text PDF
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Optimization of storage temperature for the pollen viability of transgenic plants that express the anti-breast cancer monoclonal antibody mAb BR55

Chae-Yeon Lim, Deuk-Su Kim, Kyung Jin Lee, Kyung-A Hwang, Young-Kug Choo and Kisung Ko*

Department of Medicine, Medical Research Institute, College of Medicine, Chung-Ang University, 84 Heukseok-ro, Dongjak-gu, Seoul 156-756, Korea
Department of Agrofood Resources, National Academy of Agricultural Science, RDA, Suwon, Gyeonggi-do 441-707, Korea
Division of Biological Science, College of Natural Sciences, Wonkwang University, Iksan, Jeonbuk 570-749, Korea

Abstract
Pollen germination viability is an essential factor in the production of seeds from pollination and fertilization. In this study, we optimized storage temperature conditions to increase the viability of pollen germination and pollen germ tube growth in transgenic plants that express the anti-breast cancer monoclonal antibody mAb BR55. The pollen of the transgenic plant BR55 was stored at five different temperatures: room temperature (RT), 4°C, -20°C, -70°C, or -196°C for 10 weeks. The in vitro pollen germination rate and pollen tube growth were observed at 10 weeks. The pollen was then germinated in vitro in sucrose buffer, either alone, or supplemented with 20
΅g/mL boric acid. In general, both pollen germination rate and pollen tube growth increased with boric acid supplementation, compared to the control. The pollen germination rate and the pollen germ tube length were the greatest in pollen stored at -20°C (p < 0.01). The transgenic plant carrying the anti-colorectal cancer mAb CO17-1A HC and LC genes were pollinated using pollen (mAb BR55 transgenic plant) stored at -20°C, generating F1 seedlings carrying both mAb CO17-1A and mAb BR55 HC and LC genes. These results indicate that -20°C is the optimal temperature for pollen storage and ensures the pollen germination viability of transgenic tobacco plants that express therapeutic anti-cancer mAbs.

Pages 403-409 | Full Text PDF
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Identification of QTLs for new formed root architectural traits in rice (Oryza sativa L.) after transplantation

Shukun Jiang*, Xijuan Zhang, Hui Jiang, Tongtong Wang, Guohua Ding, Shichen Sun, Liangming Bai, Fengming Zhang

Cultivation and Farming Research Institute, Postdoctoral research station, Heilongjiang Academy of Agricultural Sciences, 150086 Harbin, China

Abstract

Transplanting has been the most widespread planting technique for rice production in Asia. It is estimated that more than 85% of rice production in china, and almost 100% in Northeast China adopted transplanting. The healthy and vigorous seeding can reduce thetransplanting shock’ and is the most important for final yield formation. The large root diameter, long root length, large surface area and more root number of new formed root are the major parameters for healthy and vigorous seeding. Nipponbare, Kasalath and ninety-eight backcross recombinant inbred lines (BC1F5) derived from a cross between them were used to detect QTL controlling new formed root traits after transplanting. The healthy seeding was transplanted at fourth leaf stage. And the average root diameter, total root length, average surface area, root length and root number of new formed root were measured at 7 days after transplanting. The qARD3 associated with average root diameter was detected on chromosome 3.Two putative QTLs controlling total root length were detected on chromosomes 5 (qRL5) and 8 (qRL8).One QTL qASA5 affecting average surface area was detected on chromosomes 5. The qARL5 affecting average root length was identified at the same region with qRL5 and qASA5.Three QTLs associated with root number were detected on chromosomes 1 (qRN1), 3 (qRN3) and 5 (qRN5). The phenotypic variations (VE) explained by individual QTL were ranged from 8.6% to 24.2%. A QTLs cluster formed on chromosome 5. The qRN3 and qRL8 were new locus for new formed root traits after transplanting. The QTLs cluster on chromosome 5 was also a new region. These QTLs provided “elements” and basic-information for transplanting root architecture and elucidation of root architecture molecular mechanism after transplanting of rice.

Pages 410-414 | Full Text PDF