Frontiers of Agriculture in China >
Construction and transformation for the antisense expression vector of the polyphenol oxidase gene in Yali pear
Received date: 03 Sep 2010
Accepted date: 19 Sep 2010
Published date: 05 Mar 2011
Copyright
To inhibit the browning process in fruits of Yali pear, in this paper, antisense gene techniques were used to reduce the expression of BoldItalic gene. A cDNA fragment of 450 bp, which is located at the 3′ terminal of the polyphenol oxidase (BoldItalic) gene, was amplified from Yali pear using the RT-PCR method, then the antisense expression vector was constructed by inserting the fragment of the Yali pear BoldItalic gene between the CaMV promoter and NOS terminator of the expression vector pBI121 in a reverse orientation. After that, with the agrobacterium-mediated method, the BoldItalic antisense gene was transformed into Yali pear shoots. Northern blot analysis and enzyme activity assay showed that the PPO activities in the transgenic Yali pear shoots were significantly decreased, compared with the non-transformed Yali pear shoots. This lays a good foundation for breeding new varieties of pears with browning resistance in the future.
Guiqin LI , Jing QI , Yuxing ZHANG , Zhihua GAO , Dongqian XU , Huixuan LI , Chenmin HUO . Construction and transformation for the antisense expression vector of the polyphenol oxidase gene in Yali pear[J]. Frontiers of Agriculture in China, 2011 , 5(1) : 40 -44 . DOI: 10.1007/s11703-010-1058-y
| 1 |
Bachem C W B, Speckmann G J, van der Linde P C G, Verheggen F T M, Hunt M D, Steffens J C, Zabeau M (1994). Antisense expression of polyphenol oxidase genes inhibits enzymatic browning in potato tubers. Nat Biotechnol, 12(11): 1101–1105
|
| 2 |
Cary J W, Lax A R, Flurkey W H (1992). Cloning and characterization of cDNAs coding for Vicia faba polyphenol oxidase. Plant Mo1 Biol, 20: 245–253
|
| 3 |
Chevalier T, de Rigal D, Mbéguié-A-Mbéguié D, Gauillard F, Richard-Forget F, Fils-Lycaon B R (1999). Molecular cloning and characterization of apricot fruit polyphenol oxidase. Plant Physiol, 119(4): 1261–1270
|
| 4 |
Dong Z (2007). The study on the ACC oxidase genes clone of pear fruit. Dissertation for the Master Degree. Baoding: Agricultural University of Hebei, 84–85 (in Chinese)
|
| 5 |
Gao M, Matsuta N, Murayama H, Toyomasu T, Mitsuhashi W, Dandekar A, Tao R, Nishimura K (2007). Gene expression and ethylene production in transgenic pear (Pyrus communis cv. ‘La France’) with sense or antisense cDNA encoding ACC oxidase. Plant Sci, 173(1): 32–42
|
| 6 |
Gao M, Sakamoto A, Miura K, Murata N, Sugiura A, Tao R (2000). Transformation of Japanese persimmon (Diospyros kaki Thunb.) with a bacterial gene for choline oxidase. Molecular Breeding, 6(5): 501–510
|
| 7 |
Joy R W IV, Sugiyama M, Fukuda H, Komamine A (1995). Cloning and characterization of polyphenol oxidase cDNAs of Phytolacca americana. Plant Physiol, 107(4): 1083–1089
|
| 8 |
Li G Q, Li H X, Xu D Q, Liu K, Zhang Y X (2008). Cloning and expression of polyphenol oxidase gene of CDS from Yali pear. Journal of Fruit Science, 25(4): 577–580 (in Chinese)
|
| 9 |
Matsuda N, Gao M, Isuzugawa K, Takashina T, Nishimura K (2005). Development of an Agrobacterium-mediated transformation method for pear (Pyrus communis L.) with leaf-section and axillary shoot-meristem explants. Plant Cell Rep, 24(1): 45–51
|
| 10 |
Murashige T, Skoog F (1962). A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant, 15(3): 473–497
|
| 11 |
Newman S M, Eannetta N T, Yu H, Prince J P, Vicente M C, Tanksley S D, Steffens J C (1993). Organization of the tomato polyphenol oxidase gene family. Plant Mo1 Biol, 21: 1035–1051
|
| 12 |
Thygesen P W, Dry I B, Robinson S P (1995). Polyphenol oxidase in potato. A multigene family that exhibits differential expression patterns. Plant Physiol, 109(2): 525–531
|
| 13 |
Vámos-Vigyázó L, Haard N (1981). Polyphenol oxidase and peroxidase in fruits and vegetables. Crit Rev Food Sci Nutr, 15(1): 49–127
|
| 14 |
Van der Krol A R, Lenting P E, Veenstra J, van der Meer I M, Koes R E, Gerats A G M, Mol J N M, Stuitje A R (1988). Antisense chalcone synthrase gene in transgenetic plants inhibits flower pigmentation. Nature, 333(6176): 866–869
|
| 15 |
Vaughn K C, Lax A R, Duke S O (1988). Polyphenol oxidase: the chloroplast oxidase with no established function. Physiol Plant, 72(3): 659–665
|
| 16 |
Wang Q, Huang H Y, Ma W F, Wang D (2007). Effect of anti-sense PPO gene on the tuber-browning of Solanum tuberosum L. Acta Agron Sin, 33(11): 1822–1827 (in Chinese)
|
| 17 |
Yamamoto T, Hayasi T (2001). Mapping of peach. In: Sasaki T, Tabata S, Shimamoto K, eds. Protocol for Studying of Plant Genome. Tokyo: Shujunsha Co., Ltd., 222–225
|
/
| 〈 |
|
〉 |