Gene cloning and transformation of Arabidopsis plant to study the functions of the Early Responsive to Dehydration gene (ERD4) in coffee genome

Coffee plant is one of the most important industrial crops, and the two popular cultivars, Coffea arabica and Coffea canephora, contribute to the production of almost all coffee beans around the world. Traditional breeding could be used to develop new coffee cultivars with a higher productivity under these harsh conditions, and a biotechnological approach can also be used to improve coffee plants in a relatively short period of time.
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Gene cloning and transformation of Arabidopsis plant to study the functions of the Early Responsive to Dehydration gene (ERD4) in coffee genomeTAÏP CHÍ PHAÙT TRIEÅN KH&CN, TAÄP 19, SOÁ T3- 2016Gene cloning and transformation ofArabidopsis plant to study the functions ofthe Early Responsive to Dehydration gene(ERD4) in coffee genomeNguyen Dinh SyInstitute of Environment and Biotechnology, Taynguyen UniversityHunseung KangCollege of Agriculture and Life Sciences, Chonnam National University(Received on 23 th November 2015, accepted on May 5 th 2016)ABSTRACTCoffee plant is one of the most importantperiod of time. To develop new coffee cultivarsindustrial crops, and the two popular cultivars,via a biotechnological approach, it is necessaryCoffea arabica and Coffea canephora, contributeto discover potential candidate genes andto the production of almost all coffee beansdetermine their functions in coffee plants.around the world. Although the demand forHowever, it is technically difficult to introducecoffee beans is continually increasing, the steadyforeign genes into coffee genome and takes longproduction of coffee beans is hampered by manytime to analyze gene function in coffee plants. Tofactors, such as environmental stresses, insectovercome these technical difficulties, thepests, and diseases. Traditional breeding couldpotential coffee genes could be cloned andbe used to develop new coffee cultivars with aintroduced into Arabidopsis for the rapidhigher productivity under these harsh conditions,analysis of its biological functions under harshenvironmental conditions.and a biotechnological approach can also beused to improve coffee plants in a relatively shortKeywords: Arabidopsis, Coffee genome, gene cloning, transgenic plantINTRODUCTIONCoffee plant is a tropical crop belonging toRubiaceae family that has more than 100 specieswhich are native of African continent,Madagascar, and the Mascarene Islands [1].Although many varieties of coffee cultivars exist,most of the coffee beverages are made from twospecies, Arabica coffee (Coffea Arabica) andRobusta coffee (Coffea canephora), with exportvalues of approximately US$ 22 billion in theyear of 2012 and over 600 billion cups consumedevery year throughout the world [2]. Coffeeplants are currently cultivated in 80 countriesproducing approximately 70 % and 30 % ofArabica and Robusta beans, respectively [3]. AreportbyICO(InternationalCoffeeOrganization) indicated that ten leadingcountries, including Brazil, Vietnam, Indonesia,Colombia, Ethiopia, India, Honduras, Peru,Mexico, and Guatemala, contribute 35 %, 15.2%, 8.8 %, 7.1 %, 4.4 %, 3.7 %, 3.1 %, 3.1 %, 3.0%, and 2.6 % of world coffee bean production,respectively [2].Trang 53Science & Technology Development, Vol 19, No.T3-2016C. canephora is the diploid species (2n=22chromosomes) and is self-incompatible, whereasC. arabica is allotetraploid (2n=4x=44chromosomes) self-fertile species [4] that wasoriginated from cross between C. eugenoides andC. canephora [5]. Due to the differences inmorphological and physiological characteristics,C. canephora appears to be more vigorous,productive, and resistant to disadvantageousconditions than C. Arabica does [6]. In general,C. Arabica is preferred to C. canephora due to itslow-caffeine content and less-bitter taste.In recent years, global warming causessevere climate changes, including high and lowtemperatures, prolonged-drought season, oralteration of raining and snowing patterns, thatsignificantly affects the yield of agriculturalproducts. The productivity of coffee plants can bereduced up to 80 % by environmental stresses,including drought, salt, cold, high temperature,and UV light, especially by prolonged waterdeficiency [6]. Until now, conventional breedinghas mainly been used to improve coffee plants,but it takes a long time (approximately 30 years)and requires many steps, including selection,hybridization, and progeny evaluation, to developa new coffee cultivar via conventional breeding.Therefore, in other to develop a new coffeecultivar that has beneficial traits such as abioticand biotic stress tolerance, disease resistance, orquality and quantity improvement, more rapidand efficient strategy utilizing genetictransformation technology is required.During the last two decades, geneticresearches on coffee plants demonstrated theregulation, function, and interactions of coffeegenes. Several research groups analyzed thecoffee transcriptomes and expressed sequencetags (ESTs) from both Robusta and Arabicacoffee plants [7-8], and other groups utilizedoligo-based microarray containing 15,721unigenes to study the functions of coffee genesTrang 54involved in bean maturation or resistance topathogens or drought [9], which opens a way forfunctional genomics of coffee plants. The ESTsequences of C. arabica can be found at thepublic website (http://www.coffee.dna.net) [10],and the genome assembly and gene models of C.canephora are available on the Coffee GenomeHub (http://coffee-genome.org) [11]. In addition,transformation systems of coffee plants, utilizingelectroporation[12],microprojectilebombardment[13-17],Agrobacteriumtumefaciens [18-26], or A. rhizozenes [27-31],have been developed to deliver potential targetgenes into coffee plants. However, it takes longtime and is technically difficult to introduceforeign genes into coffee genome due to lowpercentage of successful transformation, whichsignificantly restrains the functional analysis ofpotential genes in coffee plants.To overcome these technical difficulties,more rapid and efficient system is required toanalyze the functions of coffee genes in areasonable time periods. Here, we introduce anefficient system using a model plant Arabidopsisthaliana to investigate the functions of coffeegenome, which is practical, less time- a ...