Genetic engineering of sweet potatoes (Ipomoea batatas) using isopentenyl transferase gene for enhanced drought tolerance

##plugins.themes.bootstrap3.article.main##

SYLVIA OBINDA NAWIRI
RICHARD OKOTH ODUOR
ALLAN MGUTU JALEMBA

Abstract

Abstract. Nawiri SO, Oduor RO, Jalemba AM. 2017. Genetic engineering of sweet potatoes (Ipomoea batatas) using isopentenyl transferase gene for enhanced drought tolerance. Asian J Agric 1: 85-99. Approximately 70% of yield crop reduction worldwide is caused by drought. Due to severe drought, which happens often as a result of climate change, substantial yield deprivation is usual among the major cereals such as maize, wheat, and barley. Therefore, drought tolerant crops that still yield amidst erratic climatic phenomena are greatly needed. Due to its capability to produce high yield in a short period, sweet potato is suitable for cultivation in regions with limited or erratic rainwater supply where other food crops cannot grow easily. Nevertheless, its sensitivity to water deficit may lead to adverse crop growth and yield. By conventional hybridization method, sweet potato is tried to be improved, but it gives unsatisfied results due to its high male sterility, sexual incompatibility and hexaploidy nature of its genome. The aim of this study, therefore, is to develop new varieties of sweet potato with improved tolerance to water-deficit stress for sustainable production of sweet potato under water-limited conditions. Three sweet potato genotypes: Jewel, Kemb36, and Ksp36 were transformed using isopentenyl transferase gene (IPT) that delays drought-induced senescence via up-regulation of cytokinin biosynthesis, under the control of a water-deficit responsive and maturation specific promoter (PSARK). The PNOV-IPT gene construct was introduced into sweet potatoes to evaluate their transformability and regenerability. It was done via Agrobacterium tumefaciens strain EHA101 and the plants subsequently regenerated via somatic embryogenesis. Jewel genotype recorded the highest transformation and regeneration frequency, followed by Kemb36 and KSP36. Calli were cultured on media supplemented with various mannose concentrations to evaluate the suitability of mannose as a selectable marker for sweet potato, and it was discovered that 30 g/L concentration was optimal for selection of transformed events. At the time of PCR analysis, Jewel had the highest transformation efficiency followed by Kemb36. At the time for evaluation on drought tolerance under controlled conditions, the sweet potato showed delayed senescence and greater drought tolerance under water deficit conditions in the glasshouse. These plants exhibited better growth, higher yield, higher water status maintenance, higher chlorophyll content, and thus higher photosynthetic rates under reduced water conditions in comparison to wild-type. These results, therefore, indicated that expression of isopentenyl transferase gene in sweet potato significantly improves drought tolerance. Therefore, IPT gene should be used to transform other economically important food crops to delay drought-induced senescence and enhance drought tolerance.

2017-01-01

##plugins.themes.bootstrap3.article.details##

References
Anjum SA, Wang LC, Farooq M, Hussain M, Xue LL, Zuo CM. 2011.
Brassinolide application improves the drought tolerance in maize
through modulation of enzymatic antioxidants and leaf gas exchange.
Agron Crop Sci 10: 1111-1439.

Anwar N, Watanabe KN, Watanabe JA. 2010. Transgenic sweet potato
expressing mammalian cytochrome P450. Plant Cell Tiss Organ Cult
105: 219-231.

Battisti DS, Naylor RL. 2009. Historical warnings of future food
insecurity with unprecedented seasonal heat. Science 323: 240-244.
Boyer JS. 1982. Plant productivity and environment. Science 218: 443-
448.

Cao YJ, Wei Q, Liao Y, Ling H, Li SX, Xiang CB, Kuai BK. 2009.
Ectopic overexpression of AtHDG11 in tall fescue resulted in
enhanced tolerance to drought and salt stress. Plant Cell Rep 28: 579-
588.

Chen JH, Su TC, Lin HC, Huang JG, Lin HY. 2010. Expression of sweet
potato cysteine protease SPCP2 altered developmental characteristics
and stress responses in transgenic Arabidopsis plants. J Plant Physiol
167: 838-847.

Choi TJ, Aing K, Len SI, Wu CC, Chiang SF, Su CL. 2006. Regulation of
phosphate homeostasis by micro RNA in Arabidopsis. Plant Cell 18:
412-421

Dafny-Yelin M, Tzfira T. 2007. Delivery of Multiple Transgenes to Plant
Cells. Plant Physiol 145: 1118-1128.

Ekanayake IJ. 1990. Evaluation of potato and sweet potato genotypes for
drought resistance. International Sweet potato Center (CIP) Research
Guide 19: 66-68).

FAOSTAT. 2010. Statistics databases and data-sets of the Food and
Agriculture Organization of the United Nations. FAO, Rome, Italy.
FAOUN. 2010. Food Security statistics of the World 2010. FAO, Rome,
Italy.

Gama MIC, Leite JRP, Cordeiro AR, Cantliffe DJ. 1996. Transgenic
sweet potato plants obtained by Agrobacterium tumefaciens-mediated
transformation. Plant Cell Tiss Organ Cult 46: 237-244.

George EF, Hall MA, de Klerk GJ. 2008. Plant propagation by tissue
culture. Springer, Dordrecht, Nederland.

Halliwell B. 2007. Dietary polyphenols: good, bad, or indifferent for your
health? Cardiovascular Res 73: 341-347.

Jewell CM, Bradley CC, Godwin DI. 2010. Transgenic Plants for Abiotic
Stress Resistance. Springer-Verlag, Berlin.

Jiang Q, Zhang JY, Guo X, Bedair M, Sumner L, Bouton J, Wang ZY.
2010. Improvement of drought tolerance in white clover (Trifolium
repens) by transgenic expression of a transcription factor gene WXP1.
Funct Plant Biol 37: 157-165.

Kakimoto T. 2000). identification of plant Cytokinin Biosynthesis
Enzymes as Dimethylallyl Diphosphate: ATP/ADP Isopentenyl
transferases. Plant Cell Physiol 42 (8): 677-685.

Kim YC, Ahn OY, Kim HS, Kim HY, Lee SH, Catanach SA, Jacobs
EMJ, Conner JA, Kwak S. 2010. The sweet potato IbMYB1 gene as a
potential visible marker for sweet potato intragenic vector system.
Physiologia Plantarum 139: 229-240.

Kim YH, Lim S, Yang KS, Kim CY, Kwon SY, Lee HS, Wang X, Zhou
Z, Ma D, Yun DJ, Kwak SS. 2009. Expression of Arabidopsis
NDPK2 increases antioxidant enzyme activities and enhances
tolerance to multiple environmental stresses in transgenic sweet
potato plants. Mol Breed 24: 233-244.

Kreuze JF, Klein IS, Lazaro MU, Chuquiyuri WC, Morgan GL, Mejía
PGC, Ghislain M, Valkonen JPT. 2008. RNA silencing-mediated
resistance to a crinivirus (Closteroviridae) in cultivated sweet potato
(Ipomoea batatas L.) and development of sweet potato virus disease
following coinfection with a potyvirus. Mol Plant Pathol 9: 589-598.

Lebot V. 2009. Sweet potato. In: Bradshaw JE (ed) Root and Tuber Crops.
Springer Sciences and Business Media, Dordrecht, Nederland.

Lichtenthaler HK. 1987. Chlorophyl and Caretenoids: Pigments of
photosynthetic biomembranes. Methods Enzymol 148: 350-382.

Lim PO, Kim HJ, Nam HG. 2007) Leaf senescence. Ann Rev Plant Biol
58: 115-136.

Liu X, Hua X, Guo J, Qi D, Wang L, Liu Z, Jin Z, Chen S, Liu G. 2008.
Enhanced tolerance to drought stress in transgenic tobacco plants
overexpressing VTE1 for increased tocopherol production from
Arabidopsis thaliana. Biotechnological letters 30: .1275-1280.

Long SP, Ort DR. 2010. More than taking the heat: crops and global
change. Current. Opinions. Plant Biol 13: 241-248.

Lu YY, Deng XP, Kwak SS. 2010. Over expression of CuZn superoxide
dismutase (CuZn SOD) and ascorbate peroxidase (APX) in transgenic
sweet potato enhances tolerance and recovery from drought stress.
African J Biotechnol 9: 8378-8391.

Luceri C, Giovannelli L, Pitozzi V, Toti S, Castagnini C, Routaboul J M,
Lepiniec L, Larrosa M, Dolara P. 2008. Liver and colon DNA
oxidative damage and gene expression profiles of rats fed Arabidopsis
thaliana mutant seeds containing contrasted flavonoids. Food Chem
Toxicol 46: 1213-1220.

Luo HR, Santa Maria M, Benavides J, Zhang DP, Zhang YZ, Ghislain M.
2006. Rapid genetic transformation of sweet potato (Ipomoea batatas
(L.) Lam) via organogenesis. African J Biotechnol 5: 1851-1857.

Mervat MM, El Far K, Mangoury E, Elazab HEM. 2009. Novel Plant
Regeneration for Egyptian Sweet potato (Ipomoea batatas (L.) Lam.)
Abees Cultivar via Indirect Organogenesis Stimulated by Initiation
Medium and Cytokinin Effects. Australian J Basic Appl Sci 3 (2):
543-551.

Moran R, García R, López A, Zaldua Z, Mena J, Garcia M, Armas R,
Somonte D, Rodriguez J, Gomez M, Pimentel E. 1998. Transgenic
sweet potato plants carrying the delta-endotoxin gene from Bacillus
thuringiensis var. tenebrionis. Plant Sci 139: 175-184.

Newell CA, Lowe JM, Merryweather A, Booke LM, Hamilton WDO.
1995. Transformation of sweet potato (Ipomoea batatas (L.) Lam.)
with Agrobacterium tumefaciens and regeneration of plants
expressing cowpea trypsin inhibitor and snowdrop lectin. Plant Sci
107: 215-227.

Njagi IW. 2004. Optimization of parameters for Agrobacterium-Mediated
transformation of sweet potato,

Otani M, Shimada T, Kimura T, Saito A. 1998. Transgenic plant
production from embryogenic callus of sweet potato (Ipomoea
batatas (L.) Lam.) using Agrobacterium tumefaciens. Plant
Biotechnol 15: 11-16.

Pardales J, Bañoc DM, Yamauchi A, Iijima M, Esquibel C. 2000. The
effect of fluctuations of soil moisture on root development during the
establishment phase of sweet potato. Plant Prod Sci Tokyo 3 (2): 134-
139.

Peleg Z, Reguera M, Walia H, Blumwald E. 2011. Cytokinin mediated
source-sink modifications improve drought tolerance and increase
grain yield in rice under water stress. Plant Biotechnol 9: 747-758.

Pennisi E. 2008. The blue revolution, drop by drop, gene by gene. Science
320: 171-173.

Pinheiro C, Chaves MM. 2011. Photosynthesis and drought: Can we make
metabolic connections from available data. J Exp Bot 62 (3): 869-882.

Placide R, Shimelis H, Laing M, Gahakwa D. 2013. Physiological
mechanisms and conventional breeding of sweet potato (Ipomoea
batatas (L.) Lam.) to drought-tolerance. African J Biotechnol 8 (18):
1837-1846.

Qin H, Gu Q, Zhang J, Sun L, Kuppu S. 2011. Regulated expression of an
isopentenyl transferase gene (IPT) in peanut significantly improves
drought tolerance and increases yield under field conditions. Plant
Cell Physiol 52: 1904-1914.

Rivero MR, Gimeno J, Van Deynze A, Walia H, Blumwald E. 2010.
Enhanced Cytokinin Synthesis in Tobacco Plants Expressing P SARK
: IPT Prevents the Degradation of Photosynthetic Protein Complexes
During Drought. Plant Cell Physiol 51: 1929-1941.

Rivero RM, Shulaev V, Blumwald E. 2009. Cytokinin-dependent
photorespiration and the protection of photosynthesis during water
deficit. Plant Physiol 150: 1380-1393.

Rivero RM, Kojima M, Gepstein A, Sakakibara H, Mittler R, Gepstein S,
Blumwald E. 2007. Delayed leaf senescence induces extreme drought
tolerance in a flowering plant. Proc Natl Acad Sci USA 104 (49):
19631-19636.

Santa-Maria M, Kenneth V, Pecota KV, Yencho CG, Allen G, Sosinski B.
2009. Rapid shoot regeneration in industrial ‘high starch’ sweet
potato (Ipomoea batatas L.) genotypes. Plant Cell Tiss Organ Cult
109: 109-117.

Saraswati P, Purnomo DW, Mawikere LN. 2012. The effectiveness of AM
fungal in improving the tolerance of sweet potato plants to drought
stress. International Conference on Agricultural, Environment and
Biological Sciences.

Sefasi A, Kreuze J, Ghislain M, Manrique S, Kiggundu A, Semakula G,
Mukasa S B. 2012. Induction of somatic embryogenesis in
recalcitrant sweet potato (Ipomoea batatas L.) cultivars. African J
Biotechnol 11 (94): 16055-16064.

Shinozaki K, Yamaguchi-Shinozaki K. 2007. Gene networks involved in
drought stress response and tolerance. J Exp Bot 58: 221-227.

Song GQ, Honda H, Yamaguchi K. 2004). Efficient Agrobacterium
tumefaciens-mediated transformation of sweet potato (Ipomoea
batatas (L.) Lam) from stem explants using a two-step kanamycinhygromycin selection method. In vitro Cell Dev Biol Plants 40: 359-
365.

Teow CC, Truong VD, Mcfeeters RF, Thompson RL, Pecota KV, Yencho
GC. 2007. Antioxidant activities, phenolic and ?-carotene contents of
sweet potato genotypes with varying flesh colours. J Food Chem 103:
829-838.

Umezawa T, Fujita M, Fujita Y, Yamaguchi-Shinozaki K, Shinozaki K.
2006. Engineering drought tolerance in plants: discovering and
tailoring genes to unlock the future. Curr Opin Biotechnol 17: 113-
122.

Vaccari I, Martinelli L. 2009. Evaluation of the phosphomannose
isomerase-based selection system for gene transfer in grape. Vitis 48:
137-144.

Van Heerden PDR, Laurie R. 2008. Effects of prolonged restriction in
water supply on photosynthesis, shoot development and storage root
yield in sweet potato. Physiologia Plantarum 134 (1): 99-109.

Walter A, Silk WK, Schurr U. 2009. Environmental effects on spatial and
temporal patterns of leaf and root growth. Ann Rev Plant Biol 60:
279-304.

Wang CR, Yang AF, Yue GD, Gao Q, Yin HY, Zhang JR. 2008.
Enhanced expression of phospholipase C 1 (ZmPLC1) improves
drought tolerance in transgenic maize. Planta 227: 1127-1140.

Xu ZY, Lee KH, Dong T. 2012. A vacuolar beta-glucosidase homolog
that possesses glucose conjugated abscisic acid hydrolyzing activity
play an important role in osmotic stress responses in Arabidopsis.
Plant Cell 24: 2184-2199.

Yang S, Vanderbeld B, Wan J, Huang1 Y. 2010. Narrowing down the
targets: towards successful genetic engineering of drought-tolerant
crops. Molecular Plant 3: 469-490.

Zidani S, Ferchichi A, Chaieb M. 2005. Genomic DNA extraction method
from pearl millet (Pennisetum glaucum) leaves. African J Biotechnol
4 (8): 862-866.