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Research Report A leading global R&D planning and evaluation institute for technological innovation in food, agriculture and forestry

LAST UPDATE : 2016-01-01

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Subject Studies on practical use of transgenic watermelon root-stock for coping with....
Date 2016-12-15 17:50:33 Hit 283
Contents Ⅰ. Title
Studies on practical use of transgenic watermelon root-stock for coping with climate change

Ⅱ. Research results reference
1. Applied for a patent [Transgenic plant with enhanced tolerance to heat stress and salt
stress by introducing CsGolS1 gene and use thereof] (South Korea Patent
No.10-2014-0070259).(2014. 06)
2. A technology transfer to Myungsan Seed Company, Inc. using the patent application
(South Korea Patent No.10-2014-0070259).(2015. 05)
3. Overexpression of L-type lectin-like protein kinase 1 confers pathogen resistance and
regulates salinity response in Arabidopsis thaliana (2013) Plant Science 203-204, 98-106.
4. Heterologus expression of the gourd E3 ubiquitin ligase gene LsRZF1 compromises the
drought stress tolerance in Arabidopsis thaliana (2014) Plant Physiology and Biochemistry
77, 7-14.
5. Putrescine regulating by stress-responsive MAPK cascade contributes to bacterial
pathogen defense in Arabidopsis (2013) Biochem. Biophys. Res. Commun 437, 502-508.
6. Involvement of the OsMKK4-OsMPK1 cascade and its downstream transcription factor
OsWRKY53 in the wounding response in rice (2014) Plant Pathology Journal 30, 168-177.
7. Effect of heat treatment around the fruit set region on growth and yield of watermelon
[Citrullus lanatus (Thunb.) Matsum. and Nakai](2013), Physiology and Molecular Biology of
Plants, 509-514.
8. Occurrence of Diseases and Case of Clinical Diagnosis on Watermelon in South Korea,
2008-2012 (2014), Research in Plant Disease, 8-14.
9. Effect of plant growth regulators on regeneration from the cotyledon explants in
Watermelon (Citrullus lanatus (thunb.) Matsum. & Nakai) (2014) Korean J. Plant Research
27, 51-59.
10, Several poster presentation; Transgenic watermelon root stock plants overexpressing the
cucumber galactinol synthase showed enhanced resistance against fungal pathogen. The
Korean Society for Plant Biotechnology, The lsrzf1 antisense transgenic line of the novel
Ring-type E3 ubiqutin ligase increase proline cotensts in Gong-dae watermelon cultivar.
The Korean Society for Plant Biotechnology, Cloning and expression analysis of an
arginine decarboxylase gene from bottle gourd (Lagenaria siceraria). The Korean Society
fro Plant Pathology, etc.
Ⅲ. Importance and objectives of the research
Gourd family(Lagenaria siceraria) is one of the most important agricultural crops in the
world. Water deficit is the major limiting factor for crop productivity. To improve gourd
tolerance to drought stress, identification of abiotic stress tolerance-related genes is
important. Proline accumulation is a common physiological response in higher plants
exposed to drought and salt stress [1]. Proline is considered to be an important osmolyte
that acts as a molecular chaperone to stabilize the structure of proteins, as well as a
regulator of cellular redox potential and an antioxidant controlling free radical levels [2].
Plants that are challenged by drought and salt stress recruit ABA as an endogenous signal
that initiates adaptive responses [3]. During late embryogenesis, ABA promotes the
acquisition of desiccation tolerance and seed dormancy, and inhibits seed germination [4, 5].
The molecular mechanisms underlying ABA-mediated plant tolerance to drought stress are
still not fully understood because of the complex nature of the plant response to ABA
signaling and drought stress. However, the study of stress-responsive transcription factors
has been one of the foci in studies on drought stress tolerance [6]. Protein ubiquitination is
an important post-translational modification process that is employed by eukaryotes to
regulate diverse cellular and developmental processes [7]. In higher plants, ubiquitinated
proteins are involved in abiotic or biotic stress response, hormone response, cell cycle
progression and cell differentiation [8, 9, 10, 11]. RING (for Really Interesting New Gene)
motif-containing E3 ubiquitin ligase comprises one of the largest gene families in the plant
genome. The Cys-rich RING zinc finger was first described in the early 1990s [12]. It was
defined as a linear series of conserved Cys and His residues(C3HC/HC3) that bind two
zinc atoms in a cross brace arrangement. RING zinc fingers can be divided into two types,
C3HC4 (RING-HC) and C3H2C3 (RING-H2), depending on presence of either a Cys or a
His residue in the fifth position of the motif [13]. Recently, a number of Arabidopsis RING
E3 ubiquitin ligases were shown to be involved in various cellular processes, such as auxin
signaling, ABA signaling, brassinosteroid response, seed germination, seedling development,
adaptive pathway to nitrogen limitation, and sugar responses [9, 14, 15, 16, 17]. In
particular, RING proteins play a key role in the response to environmental stimuli. For
example, they participate in photomorphogenesis, defense signaling, senescence, and
tolerance mechanisms against cold, drought, salt and osmotic stress [6, 8, 18, 19]. Recently,
we described the functional characterization of a C3H2C3-type RING-H2 zinc finger
protein, designated Arabidopsis thaliana RING Zinc Finger 1(AtRZF1) [20]. Functional
studies demonstrated that AtRZF1 participates negatively in proline production under
drought condition. Water deficit response assays indicated that, while the atrzf1 mutant
was less sensitive to drought, AtRZF1-overexpressing plants were more sensitive,
suggesting that AtRZF1 negatively regulates the drought response during early seedling
development. Furthermore, our previous data reported that AtRZF1 is a functional ubiquitin
E3 ligase [20]. In this research, an ortholog of the AtRZF1 gene in the gourd species
Lagenaria siceraria, LsRZF1, was cloned to examine the role of LsRZF1 by expression in
Arabidopsis. The gene was reduced in response to drought stress and ABA phytohormone.
The response of constitutive expression, leading to both complementation and ectopic
expression of LsRZF1 in the atrzf1 mutant and transgenic Arabidopsis plants, respectively,
against different abiotic stresses was studied. LsRZF1-ectopic expressing plants were
hypersensitive to osmotic stress in terms of cotyledon development due to the reduction in
the contents of proline and regulated the expression of stress-related genes. The
physiological processes of the drought induced phenotype began faster in the
LsRZF1-ectopic expressing lines than in the wild type(WT) and atrzf1 complementation
plants. Additionally, ubiquitin E3 ligase activity and genetic data indicated that AtRZF1 and
LsRZF1 function in a similar pathway to control proline metabolism in Arabidopsis under
drought condition. Bottle gourd has been used as a source of rootstock for watermelon
against soil-borne disease and low soil temperature. However, the wilt of cucurbit crops in
the field was caused by abiotic stress such as drought. The improved stress tolerance of
bottle gourd rootstock will be needed for breeding program. Therefore, we tried to clone an
arginine decarboxylase(ADC) gene, which is involved in plant putrescine(Put) biosynthesis,
from bottle gourd and to analyze its expression in abiotic stress conditions. Transcripts of
LsADC and LsSAMDC in bottle gourd leaves were induced continuously in response to
drought and high salt treatment and pathogen infection as well. Taken together, LsADC
and LsSAMDC are stress-responsive genes and could be used as candidate genes for
bottle gourd genetic transformation in the future. In addition, polyamines in plants are
involved in various physiological and developmental processes including abiotic and biotic
stress responses [21,22]. We investigated the expression of ADCs, which are key enzymes
in putrescine(Put) biosynthesis, and roles of Put involving defense response in Arabidopsis.
Finally, the disease susceptibility of the adc2 mutant was recovered by the addition of
exogenous Put [23,24]. Also, we were able to measure putrescine accumulation in
transgenic Arabidopsis plants harboring LsADC gene compared to wild type. The
transgenic LsADC Arabidopsis plants confers tolerance to drought stress. These results
suggest that Put is believed to have to have an osmoprotectant function in plants cells
inder water deficit. Management of irrigation in greenhouse is one of environmental factors
that affect on watermelon growth and fruit quality. Moreover, soil moisture provides
favorable environment for pathogens. Zoospore of olpidium moves to another host and
reproduce through water and it can be a vector of Melon necrotic spot virus(MNSV). The
occurrence of MNSV is mostly observed in high soil moisture. Infected plants showed poor
growth led to reproduction of small fruits and sugar content and value of commodities
were decreased. Thus, the management of irrigation in greenhouse can reduce the
occurrence of fungal diseases as well as virus vectors. The utilization of drought tolerance
cultivar is one of methods to decrease water supply. Guard rootstocks have been used for
grafting of watermelon are sensitive to dry and high moisture. Therefore, the development
of new rootstocks is needed. Since MNSV has been found on watermelon that use guard
as rootstocks, the development of rootstocks resistant to MNSV is considered to be crucial.



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