Day 2 :
University of Missouri, USA
Keynote: RNA-Seq Analysis Reveals Host Plant Transcriptomes in Response to Agrobacterium-mediated Transformation
Time : 09:45-10:35
Zhanyuan J. Zhang has expertise in plant genetic transformation and gene regulation. He got his Ph.D. in the area of plant genetic engineering at University of Nebraska-Lincoln, NE, USA. He has been the director of Plant Transformation Core Facility at University of Missouri, Columbia, Mo, USA since 2000. The mission of his core facility is to enhance both basic and applied plant biology studies by providing plant transformation services and to advance transgenic technologies. He has contributed to the transformation system improvements in soybean, maize, sorghum, wheat, and switchgrass. His interest in gene regulation has led to the validation and revealing of effective RNAi in soybean and novel approach for plant gene silencing employing artificial trans-acting siRNA. His research efforts in basic study of Agrobacterium-mediated transformation has discovered the role pf the heat shock protein 90.1 in this T-DNA transfer process.
Agrobacterium-mediated plant transformation has become a predominant tool for many basic studies and biotechnological applications. Discoveries in molecular mechanisms governing this transformation process have significant implications in both basic and applied plant biotechnological applications. To date, however, knowledge about plant genes and associated pathways involved in the Agrobacterium-mediated T-DNA transfer has been very limited. Here, we employed RNA-seq to exploit Arabidopsis thaliana transcriptomes in responses to Agrobacterium transformation process. We used two contrasting Agrobacterium strains to infect Arabidopsis young seedlings using AGROBEST assay protocol. The two strains included a non-oncogenic disarmed Agrobacterium strain, A136, and At804 which is a derivative of EHA105 and contains a disarmed super virulent Ti-plasmid and a binary vector pBISN1. The strain A136 lacks Ti-plasmid and therefore is unable to deliver T-DNA and effector (Vir) proteins to plant cells. This is in contrast to At804 which is capable of transferring both T-DNA and effector proteins into plant cells. Arabidopsis tissue samples for RNA-Seq were from three different treatment conditions, i.e., mock, A136 and At804, at 6 different time points (0, 3, 6, 12, 24, and 48 hours), respectively, during the Agrobacterium infection. Total RNA samples at each time point were then subject to NGS analysis. The transcriptomic analysis results showed that many plant genes responded to Agrobacterium infection. GO (gene ontology) analysis revealed that many plant biological processes are involved during Agrobacterium-plant interactions. These processes include hormone signaling, defense response, cellular biosynthesis, and nucleic acid metabolism and so on. Key genes displaying substantial changes in their transcripts were further validated by qRT-PCR and mutant screen. More details will be presented.
Universiti Kebangsaan Malaysia, Malaysia
Time : 10:35-11:25
Roohaida Othman received her PhD in Biochemistry from University of Southampton, and joined Universiti Kebangsaan Malaysia as a lecturer immediately after in 1995. Her research interest is focused on understanding the molecular mechanisms underlying the biosynthesis of commercially important metabolites in plants and algae. Her research group has employed tools of molecular biology, biochemistry and physiology as well as genomics and transgenics technology platforms to study the enzymes involved in these pathways. They have also developed protocols for higher plant and algae RNA extraction methods and overexpression of recombinant proteins in bacterial systems. She has been Editor-in-Chief for the Journal of Tropical Plant Physiology since 2010 and has been reviewer for several journals including International Journal of Food Properties.
Statement of the Problem: Persicaria minor (synonym Polygonum minus). produces a broad range of secondary metabolites such as sesquiterpenes that contribute towards the unique aroma of this plant. In an effort to understand the biosynthesis of these compounds, candidate genes involved in the sesquiterpene biosynthetic pathway have been identified from an expressed sequences tags (ESTs) collection. The purpose of this study is to characterize a gene which was initially identified as an-alpha farnesene synthase gene from the EST studies. Methodology & Theoretical Orientation: The full length cDNAs from P. minus was isolated and cloned into Escherichia coli, Lactococcus lactis and Arabidopsis thaliana following standard protocols. Enzymatic assay for the recombinant sesquiterpene synthase was performed using farnesyl diphosphate as substrate and the products from the enzymatic assays were analyzed using gas chromatography-mass spectrometry (GC-MS). Findings: The full length sequence of P. minor sesquiterpene synthase cDNA clone (PmSTS) was 2035 bp in size and was expressed in E. coli as a ~65 kDa soluble protein whereas in L. lactis, the size of the recombinant protein was ~63kDa. For enzyme activity assay, the major product of the recombinant PmSTS in E. coli was α-farnesene whilst for L. lactis recombinant protein, the major product was 𝛽-sesquiphellandrene with beta-farnesene as a minor product. Subsequent expression in A. thaliana also produced transgenic lines with increased 𝛽-sesquiphellandrene production. Finally, new expression in E. coli produced a recombinant PmSTS that released 𝛽-sesquiphellandrene as a major product and 𝛽-farnesene as a minor product, similar to the L. lactis recombinant protein. Conclusion & Significance: The identity of a plant sesquiterpene synthase has been confirmed as a 𝛽-sesquiphellandrene synthase. The correct identity of the gene was finally achieved due to the updated version of the mass spectral library used in identifying the products from GC-MS.
- Traditional medicine | Plant genetics and Plant genomics
Location: Holiday Inn Bangkok
University of Wyoming, USA
Title: Ethno-medicinal plant diversity of Bangladesh – their conservation status at the botanical garden of Bangladesh Agricultural University
Time : 11:45-12:20
AKM Golam Sarwar, a Professor of Plant Systematics at the Bangladesh Agricultural University, has his expertise in Bio-resources and Production Science (major in Pollen Morphology and Ericaceae Systematics). Four discoveries in angiosperm palynology and another one in seed physiology of rice. Recipient of “Young Researcher Award 2009” of Palynological Society of Japan. Graduate student adv isory committee Chair/Member: MS 5 (ongoing), 17 (Completed) and PhD 5 (ongoing). Now, Dr Sarwar is leading huge conservation activities on ethno-medicinal plant biodiversity of Bangladesh as the Curator, Botanical Garden, the 2nd largest of this type in Bangladesh, of Bangladesh Agricultural University. His current research interests are on Ethno-botany, Plant Morphology, Biodiversity and Conservation related issues, and Bio-energy crops.
Statement of the Problem: Ethno-Medicinal plant genetic resources are one of the most important elements of biodiversity which support life system on earth. Over 50% of prescription drugs are derived from chemicals those first identified in plants. Bangladesh, as a part of the ancient Indian subcontinent, has a long history on use of plants in the traditional medicine as Ayurvedic, Unani and Tibetan System of Medicine. Although occupies relatively small geographical area, She is rich in both floral and faunal diversities. Bangladesh is also rich in ethnic minority population and people of more than 27 ethnic minorities groups live in Bangladesh. The flowering plants diversity in Bangladesh varies from 3,813 to 5,700 species. More than 750 species have been prescribed for the treatment of different diseases in traditional medicine. The Botanic Gardens Conservation International identified 400 medicinal plants at risk of world-wide extinction from over-collection and deforestation, threatening the discovery of future cures for disease. On contrary, an estimated 80% of people, the majority of these people in developing countries, worldwide rely on traditional, largely herbal, medicine to meet their primary healthcare needs. On this back ground, the Botanical garden of Bangladesh Agricultural University has initiated programmes on ex situ conservation of plants with ethno-medicinal importance along with other conservation activities from its inception in 1963.Findings & Conclusion: The Botanical garden has harbored a large collection of ethno-medicinal plants; more than 350 species have been conserved, and the number is ever increasing. Among these, 23 species are threatened in Bangladesh territory; however, many of them are rare in the wild. The important uses of collected medicinal plants have also been described. As the population density is increasing in an alarming rate, proper attention should be given to conserve the ethno-medicinal plant resources of Bangladesh for the welfare of human being and animal health.
Assam Agricultural University, India
Title: How wheat (Triticum aestivum L.) crop does respond to aerosols of oxidized and reduced nitrogen along with light regime, physiological drought and substratum types?
Time : 12:20-12:55
Academic degrees: B.Sc.(Agri,), M.Sc.(Agri), Ph.D.(Biology), DIC (Bryophyte Physiology) London University. At present position is working as a Professor & Head (Crop Physiology), Assam, Agricultural University, India Hobby: Travelling, Reading, Playing with kids and cooking at home Challenges: Meeting Deadlines, Fighting with Destiny etc.
The aim of the investigation was to study the impacts of some abiotic factors on the responses of wheat crop (variety: Ankur Omkar) to oxidised (NaNO2) and reduced (NH4Cl) aerosols in field situation. Natural light was reduced to ≈50% using standard hessian cloth, and physiological drought was induced by spraying PEG-6000 @5000ppm (≈0.05bar). Substratum types were natural acid soil with FYM @5 t/ha (pH 5.03), acid-mineral soil (natural acid soil with FYM @10 t/ha, pH between 5.92) and acid-mineral soil mixed with FYM @10 t/ha added with lime @ 0.5t/ha (pH 6.46).There were positive effects of the aerosols on wheat crop under normal light condition, and low light suppressed the nitrogen assimilation and physiological performance of wheat. During the physiological drought condition, the aerosols showed negative impacts on wheat crop. The reduced aerosol acted as an acidifying agent, which was deleterious to the crop. Soil amended with higher dose (10t/ha) of FYM further exerted negative influence to the crop. Acid soil amended with lime (supply of Ca2+) ameliorates the negative impacts of the aerosols on yield attributes of wheat crop.
Statement of the Problem: Atmospheric wet and dry depositions of Nitrogen are important processes in the redistribution of nitrogen throughout the environment. Nitrogen oxides (NO2, NO but not N2O) reacting with intercellular water get converted into HNO2 and HNO3, which then dissociate to form nitrate, nitrite and protons. Higher concentration (<10 µll-1) of oxides of nitrogen alter the physiological processes including net photosynthesis, dark respiration, root:shoot ratios in plants and yields. Cellular plasmolysis is caused by the lipid breakdown in membrane. Acidification of the ecosystem may also result from the deposition of gaseous NH3 and particulate NH4+ (collectively NHy). Plants fed with Ammonia at high concentration (>1mM) suffer from its toxicity.
Saraswathi Narayanan College, India
Title: Impact of Neem and Pongamia oil formulation and plant extracts for the management of berry borers of organic coffee plants as part of coffee integrated pest management
Time : 13:55-14:30
He has been involved in Coffee Production both organic and conventional for the last 7 years in the Western Ghats of Palni hills. At present, a Doctoral course student in the Department of Botany, Saraswathi Narayanan College (Autonomous) Perungudi, Madurai, Tamil Nadu, India. Recently, He won the second prize in International Conference. He has served as a Manager in two Coffee Estates and modernized the estates. His interest and commitment to ecology and plant science began in 1990. As a under graduate he published his first Biotechnology paper on Plant Regeneration through somatic embryogenesis from mature leaf explants of Eryngium Foetidum, a condiment in plant cell, Tissue and Organ Culture 56: 131-137, 1999, Netherlands. Earlier he also has served as a Principal and Correspondent of St. Joseph’s Industrial School an ISO9000-2000 certified Institution, Ooty for five years. As a Philanthropist and Quality manager he has qualified himself in MBA Personnel Management and diploma in Magnetic therapy and Acupressure
Three consecutive years’ experiments were conducted at the organic coffee fields of Loyola Estate, Sirumalai near Dindigul District of Tamil Nadu, South India during 2013, 14 and 15 to evaluate bioefficiency of plant oils and leaf, garlic extracts against berry borers in the chosen organic coffee field. Among these plant oils tested, spot application of Neem and pongamia seed oils along with emulsifier was found to be very effective in the initial stage causing 90 to 94 percent reduction in borer population over control after three rounds of spraying in the final year. The overall borer population was found to be 90 percent reduced in the coffee plants sprayed with mixture of Neem with pongamia oil 3 percent and 2 percent of garlic extract even at the later stage of borer attack. This was followed by strict regulation of shade as per integrated pest management techniques. The percentage of damaged green fruits in the first year were 36 percent followed by 22 percent in the second year and 14 percent in the third year in treated plants against control 100%. Further, the mean grade index recorded at the time of harvest was also very low which statistically different from chemically treated plants. The other plant oil formulation viz., Pongamia and Iluppai were less effective and the leaf extract. Ipomea, Jatropha, Neem, Acorus and Chilli were not effective.
National Bank of Genoa, Tunisia
Title: Description and analysis of phenotypic diversity in Daucus L. germplasm collection in Tunisia for species and subspecies identification
Time : 14:30-15:05
Najla Mezghani is an assistant Professor in the National Gene Bank of Tunisia. She has her expertise in plant biotechnology and genetics. She is working in the field of plant genetic resources and she is particularly responsible of the ‘Vegetable, condiment and ornamental genetic resources conservation and evaluation’ program.
Statement of the problem: Daucus carota L. is a morphologically diverse species found throughout the Mediterranean regions and in many continents worldwide. Among Mediterranean regions, Tunisia is considered a center of biodiversity for Daucus and many other crops because of the diverse ecosystems and climatic conditions. Although some floristic treatments have been published in the past few decades, many of Daucus species are without an adequate description. The purpose of the present study is to analyze the patterns of phenotypic diversity in a Tunisian Daucus collection in order to elucidate the interrelationship between the conserved accessions and to verify the suitability of morphological characterization for species and subspecies identification in our collection. Plant material and methodology: A total of 120 Daucus accessions including cultivated carrot (D. carota subsp. sativus) and wild relatives from different geographic and bioclimatic regions in Tunisia were surveyed and characterized morphologically using 30 qualitative parameters related to vegetative and reproductive parts of the plant. Quantiﬁcation of variability for each character was investigated using the standardized Shannon–Weaver diversity index (H’). Diversity was established by multiple correspondence analysis and cluster analysis. Findings: The estimated H’ index ranged from monomorphic for umbel type and position of involucral bracts on primary umbel to highly polymorphic for other traits. The highest (0.99) and the lowest (0.24) H’ values were recorded for flowering pattern within plants and foliage coverage traits respectively. Multivariate analysis and cluster analysis permitted the subdivision of the Daucus collection into 9 distinct groups supporting traditional taxonomic treatments with a distinction of cultivated carrot from the closely related wild species. Conclusion: Morphological data provide considerable information that is useful to distinguish species and subspecies in the difficult Daucus genus. Our results serve as a basis for verification and possible reidentification of Daucus accessions in Tunisia and elsewhere.
The University of Queensland, Australia
Title: Biochemical and functional characteristic of Davidsonia pruriens and Davidsonia jerseyana fruit tea
Time : 15:05- 15:40
Nilesh Nirmal has his expertise in evaluation of plant based food additive as replacement to synthetic additives. He has extensive experience in phytochemical analyses, various antioxidant assay, antimicrobial assay, anti-denaturation assay, antimelanotic assay. He also has interest in enzyme purification, characterization and its application. He introduced the low cost simple one step preparation of brazilin rich compound through column chromatography (Nirmal and Panichayupakaranant, 2014). This technique reduces the extensive financial burden of purification industry. He had developed natural plant based additives for inhibition of blackening in prawn (shrimp). This approach led to the green plant based additives which ultimately help food industries as well as consumer acceptance and well-being.
Statement of the Problem: Davidson’s plum (Davidsonia pruriens and Davidsonia jerseyana) is one of the Australian native fruits originated from the North Queensland. The present study was conducted to evaluate and compare the biochemical and functional characteristics of Davidsonia pruriens (DP) and Davidsonia jerseyana (DJ) fruit tea. Moreover, sensory analyses of fruit teas were also conducted to examine the consumer acceptance. Methodology: Dried fruits sheets were brewed with boiled water (2g / 250ml) for 5 min and extract collected. Samples were subjected to biochemical analyses. The phenolic compounds in the samples were assessed by using UHPLC system (Thermo Scientific, Waltham, MA) coupled with MS. Chromatographic separation was carried out with mobile phase A (H20 containing 0.1% formic acid) and mobile phase B (acetonitrile containing 0.1% formic acid). Organic acid in the samples were analyzed using a binary HPLC pump with photodiode array detector. The antioxidant capacity of samples was determined by using DPPH radical scavenging activity. Sensory evaluation of samples was conducted by using 9-point hedonic scale. Findings: Total phenolic content in DP and DJ was 10.37 and 11.32 mg GAE/g of dry sheet, respectively. UPHLC analysis of DP and DJ indicated gallic acid was the major phenolic compound with DJ having significantly higher level (P<0.05). Organic acid analysis showed the presence of only mallic acid in both samples. Antioxidant activities of DP and DJ were comparable (P>0.05). Flavor and taste score was higher for DJ compared to DP (P<0.05). However, overall acceptance score for both samples was not significantly different. Conclusion & Significance: DJ contained high level of gallic acid as well as mallic acid as compared to DP, which affect the taste of the DP fruit tea. However, likeness score for Davidsonia plum fruit showed promising results for Davidsonia pruriens and Davidsonia jerseyana as a fruit tea.