Please wait a minute...
Journal of Integrative Agriculture  2016, Vol. 15 Issue (2): 282-294    DOI: 10.1016/S2095-3119(15)61075-9
Crop Genetics · Breeding · Germplasm Resources Advanced Online Publication | Current Issue | Archive | Adv Search |
Cloning, localization and expression analysis of two fw2.2-like genes in small- and large-fruited pear species
 TIAN Jia, ZENG Bin, LUO Shu-ping, LI Xiu-gen, WU Bin, LI Jiang
1、College of Forestry & Horticulture, Xinjiang Agricultural University, Urumqi 830052, P.R.China
2、College of Agronomy, Xinjiang Agricultural University, Urumqi 830052, P.R.China
3、Zhengzhou Fruit Research Institute, Chinese Academy of Agricultural Sciences, Zhengzhou 450009, P.R.China
4、Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing
100193, P.R.China
Download:  PDF in ScienceDirect  
Export:  BibTeX | EndNote (RIS)      
摘要  Fruit size is one of the most important agronomic characters, which is mainly determined by cell number and cell size. However, our current knowledge about pear is largely unknown. Through counting of pear mesocarp cells at different stages, we found that the cell number, rather than the cell size, is responsible for the differences between small- and large-fruited cultivars. Fruit weight-2.2 (fw2.2) is an important quantitative trait locus (QTL) affecting fruit weight in tomato and functions as a negative regulator in carpel cell division. To get more insights into this QTL in pear fruit development, we isolated two putative homologous fw2.2 genes, which were designated as fw2.2-like (PbFWL) genes. PbFWLs encode Cys-rich proteins with the CCXXXXCPC motif and belong to the PLAC8 superfamily. In addition, results from the subcellular localization indicated that PbFWLs were localized in the plasma membrane. The expression profile of the PbFWL genes by qRT-PCR showed they expressed higher in small-sized fruit cultivar than that in large-sized fruit cultivar during the cell division period. In summary, our data suggest that these two PbFWLs might be negatively related to the cell division in pear fruit.

Abstract  Fruit size is one of the most important agronomic characters, which is mainly determined by cell number and cell size. However, our current knowledge about pear is largely unknown. Through counting of pear mesocarp cells at different stages, we found that the cell number, rather than the cell size, is responsible for the differences between small- and large-fruited cultivars. Fruit weight-2.2 (fw2.2) is an important quantitative trait locus (QTL) affecting fruit weight in tomato and functions as a negative regulator in carpel cell division. To get more insights into this QTL in pear fruit development, we isolated two putative homologous fw2.2 genes, which were designated as fw2.2-like (PbFWL) genes. PbFWLs encode Cys-rich proteins with the CCXXXXCPC motif and belong to the PLAC8 superfamily. In addition, results from the subcellular localization indicated that PbFWLs were localized in the plasma membrane. The expression profile of the PbFWL genes by qRT-PCR showed they expressed higher in small-sized fruit cultivar than that in large-sized fruit cultivar during the cell division period. In summary, our data suggest that these two PbFWLs might be negatively related to the cell division in pear fruit.
Keywords:  pear       fruit size       fw2.2       subcellular localization       gene expression       cell division  
Received: 06 January 2015   Accepted:
Fund: 

This work was supported by the National High-Tech Research and Development Program of China (2011AA10020604), the Pomology Major Projects of Xinjiang Uygur Autonomous Region of China (201130102–1) and the Special Research Projects of National Forestry Industry of Public Benefit, China (201304701–1).

Corresponding Authors:  LI Jiang,Tel/Fax: +86-991-8762363, E-mail: lijiangxnd@163.com;WU Bin, Tel: +86-10-57833201, E-mail: bwu@implad.ac.cn   
About author:  TIAN Jia, E-mail: terrisay@163.com;

Cite this article: 

TIAN Jia, ZENG Bin, LUO Shu-ping, LI Xiu-gen, WU Bin, LI Jiang. 2016. Cloning, localization and expression analysis of two fw2.2-like genes in small- and large-fruited pear species. Journal of Integrative Agriculture, 15(2): 282-294.

Al-Hina Y K, Rope T R. 2004. Rootstock effects on growth,cell number, and cell size of ‘Gala’ apples. Journal of theAmerican Society for Horticultural Science, 129, 37-41

Alpert K B, Grandillo S, Tanksley S D 1995 fw 2.2: A majorQTL controlling fruit weight is common to both red- andgreen-fruited tomato species. Theoretical and AppliedGenetics, 91, 994-1000

Altschul S F, Madden T L, Schaffer A A, Zhang J, Zhang Z, MillerW, Lipman D J. 1997. Gapped BLAST and PSI-BLAST:A new generation of protein database search programs.Nucleic Acids Research, 25, 3389-3402

Cong B, Liu J, Tanksley S D. 2002. Natural alleles at a tomatofruit size quantitative trait locus differ by heterochronicregulatory mutations. Proceedings of the NationalAcademy of Sciences of the United States of America, 99,13606-13611

Cong B, Tanksley S D. 2006. FW2.2 and cell cycle control indeveloping tomato fruit: A possible example of gene cooptionin the evolution of a novel organ. Plant MolecularBiology, 62, 867-880

Cowan A, Cripps R, Richings E, Taylor N. 2001. Fruit size:Towards an understanding of the metabolic control of fruitgrowth using avocado as a model system. PhysiologiaPlantarum, 111, 127-136

Dahan Y, Rosenfeld R, Zadiranov V, Irihimovitch V. 2010. Aproposed conserved role for an avocado FW2.2-like geneas a negative regulator of fruit cell division. Planta, 232,663-676

Espunya M C, Combettes B, Dot J, Chaubet-Gigot N, MartinezM C. 1999. Cell-cycle modulation of CK2 activity in tobaccoBY-2 cells. The Plant Journal, 19, 655-666

Fan C, Xing Y, Mao H, Lu T, Han B, Xu C, Li X, Zhang Q.2006. GS3, a major QTL for grain length and weight andminor QTL for grain width and thickness in rice, encodes aputative transmembrane protein. Theoretical and AppliedGenetics, 112, 1164-1171

De Franceschi P, Stegmeir T, Cabrera A, van der Knaap E,Rosyara U R, Sebolt A M, Dondini L, Dirlewanger E, Quero-Garcia J, Campoy J A, Iezzoni A F. 2013. Cell numberregulator genes in provide candidate genes for the controlof fruit size in sweet and sour cherry. Molecular Breeding,32, 311-326

Frary A, Nesbitt T C, Grandillo S, Knaap E, Cong B, Liu J,Meller J, Elber R, Alpert K B, Tanksley S D. 2000. fw2.2:Aquantitative trait locus key to the evolution of tomato fruitsize. Science, 289, 85-88

Gan L, Tang H R, Dong X L. 2006. An overview on identification of pear species and cultivars. Chinese Agricultural ScienceBulletin, 22, 302-307 (in Chinese)

Gehrig H H, Winter K, Cushman J, Borland A, Taybi T. 2000. Animproved RNA isolation method for succulent plant speciesrich in polyphenols and polysaccharides. Plant MolecularBiology Reporter, 18, 369-376

Grandillo S, Ku H, Tanksley S. 1999. Identifying the lociresponsible for natural variation in fruit size and shape intomato. Theoretical and Applied Genetics, 99, 978-987

Guo M, Rupe M A, Dieter J A, Zou J, Spielbauer D, DuncanK E, Howard R J, Hou Z, Simmons C R. 2010. CellNumber Regulator1 affects plant and organ size in maize:Implications for crop yield enhancement and heterosis. ThePlant Cell, 22, 1057-1073

Harada T, Kurahashi W, Yanai M, Satoh T. 2005. Involvement ofcell proliferation and cell enlargement in increasing the fruitsize of Malus species. Scientia Horticulturae, 105, 447-456

Hu Y, Xie Q, Chua N H. 2003. The Arabidopsis auxin-induciblegene ARGOS controls lateral organ size. The Plant Cell,15, 1951-1961

Kajiura I, Sato Y. 1990. Recent progress in Japanese pear(Pyrus pyrifolia Nakai) breeding and descriptions of cultivarsbased on literature review. Bulletin of Fruit Trees ResearchStation Extra, 1, 1-329

Kenis K, Keulemans J, Davey M W. 2008. Identification andstability of QTLs for fruit quality traits in apple. Tree Geneticsand Genomes, 4, 647-661

Kortstee A J, Appeldoorn N J, Oortwijn M E, Visser R G. 2007.Differences in regulation of carbohydrate metabolism duringearly fruit development between domesticated tomato andtwo wild relatives. Planta, 226, 929-939

Krizek B A. 1999. Ectopic expression of AINTEGUMENTAin Arabidopsis plants results in increased growth of floralorgans. Developmental Genetics, 25, 224-236

Li X G, Yang J, Wang L, Wang S K. 2010. Progress andproposal in research of pear breeding in China. Journal ofFruit Science, 27, 987-994 (in Chinese)

Libault M, Stacey G. 2010. Evolution of FW2.2-like (FWL) andPLAC8 genes in eukaryotes. Plant Signaling & Behavior5, 1226-1228

Libault M, Zhang X C, Govindarajulu M, Qiu J, Ong Y T,Brechenmacher L, Berg R H, Hurley-Sommer A, Taylor CG, Stacey G. 2010. A member of the highly conserved FWL(tomato FW2.2-like) gene family is essential for soybeannodule organogenesis. The Plant Journal, 62, 852-864

Liebhard R, Kellerhals M, Pfammatter W, Jertmini M, GesslerC. 2003. Mapping quantitative physiological traits in apple(Malus x domestica Borkh.). Plant Molecular Biology, 52,511-526

Liu J, Cong B, Tanksley S D. 2003. Generation and analysisof an artificial gene dosage series in tomato to study themechanisms by which the cloned quantitative trait locusfw2.2 controls fruit size. Plant Physiology, 132, 292-299

Livak K J, Schmittgen T D. 2001. Analysis of relative geneexpression data using real-time quantitative PCR and the2-ΔΔCt method. Methods, 25, 402-408

Marchler-Bauer A, Anderson J B, Cherukuri P F, DeWeese-Scott C, Geer L Y, Gwadz M, He S, Hurwitz D I, Jackson JD, Ke Z, Lanczycki C J, Liebert C A, Liu C, Lu F, MarchlerG H, Mullokandov M, Shoemaker B A, Simonyan V, SongJ S, Thiessen P A, et al. 2005. CDD: A Conserved DomainDatabase for protein classification. Nucleic Acids Research,33, D192-196

Mizukami Y, Fischer R L. 2000. Plant organ size control:AINTEGUMENTA regulates growth and cell numbers duringorganogenesis. Proceedings of the National Academy ofSciences of the United States of America, 97, 942-947

Moreno-Romero J, Espunya M C, Platara M, Arino J, Martinez MC. 2008. A role for protein kinase CK2 in plant development:Evidence obtained using a dominant-negative mutant. ThePlant Journal, 55, 118-130

Scorza R, May L, Purnell B, Upchurch B. 1991. Differences innumber and area of mesocarp cells between small-fruitedand large-fruited peach cultivars. Journal of the AmericanSociety for Horticultural Science, 116, 861-864

Shomura A, Izawa T, Ebana K, Ebitani T, Kanegae H, KonishiS, Yano M. 2008. Deletion in a gene associated with grainsize increased yields during rice domestication. NatureGenetics, 40, 1023-1028

Song W Y, Hortensteiner S, Tomioka R, Lee Y, Martinoia E.2011. Common functions or only phylogenetically related?The large family of PLAC8 motif-containing/PCR genes.Molecules and Cells, 31, 1-7

Song X J, Huang W, Shi M, Zhu M Z, Lin H X. 2007. A QTLfor rice grain width and weight encodes a previouslyunknown RING-type E3 ubiquitin ligase. Nature Genetics,39, 623-630

Sun W, Cao Z Y, Li Y, Zhao Y X, Zhang H. 2007. A simple andeffective method for protein subcellular localization usingAgrobacterium-mediated transformation of onion epidermalcells. Biologia, 65, 529-532

Tamura K, Dudley J, Nei M, Kumar S. 2007. MEGA4: MolecularEvolutionary Genetics Analysis (MEGA) software version4.0. Molecular Biology and Evolution, 24, 1596-1599

Tanksley S D. 2004. The genetic, developmental, and molecularbases of fruit size and shape variation in tomato. The PlantCell, 16(Suppl.), S181-S189.Westwood M, Batjer L, Billingsley H. 1967. Cell size, cellnumber, and fruit density of apples as related to fruit size,position in cluster, and thinning method. Proceedings ofthe American Society for Horticultural Science, 9, 151-162

Wu J, Wang Z, Shi Z, Zhang S, Ming R, Zhu S, Khan M A, TaoS, Korban S S, Wang H, Chen N J, Nishio T, Xu X, CongL, Qi K, Huang X, Wang Y, Zhao X, Deng C, Gou C, et al.2013. The genome of the pear (Pyrus bretschneideri Rehd.).Genome Research, 23, 396-408

Wu T, Tao S T, Zhang H P, Song Y, Yao G F, Zhang S L. 2011.Effects of fruit thinning on fruit sugar accumulation and leafphotosynthetic characteristics of pear. Acta HorticulturaeSinica, 38, 2041-2048 (in Chinese)

Xie H, Gao Q M, Liu Y J, Nan X, Zhang W, Li S Q, Li J. 2013.Effects of xenia on fruit quanlity of Korla fragrant pearcultivar. Acta Agriculturae Boreali-occidentalis Sinica, 22,93-96 (in Chinese)

Xu K, Huang X, Wu M, Wang Y, Chang Y, Liu K, Zhang J, ZhangY, Zhang F, Yi L, Li T, Wang R, Tan G, Li C. 2014. A rapid,highly efficient and economical method of Agrobacteriummediatedin planta transient transformation in living onionepidermis. PLOS ONE, 9, e83556.

Zhang C, Tanabe K, Wang S, Tamura F, Yoshida A, MatsumotoK. 2006. The impact of cell division and cell enlargementon the evolution of fruit size in Pyrus pyrifolia. Annals ofBotany, 98, 537-543

Zhang C Z, Ma X T. 1995. Comparison of several commonmethods to measure the volume of fruit. China Fruits, 2,44-45 (in Chinese)
[1] Qing Li, Zhuangzhuang Sun, Zihan Jing, Xiao Wang, Chuan Zhong, Wenliang Wan, Maguje Masa Malko, Linfeng Xu, Zhaofeng Li, Qin Zhou, Jian Cai, Yingxin Zhong, Mei Huang, Dong Jiang. Time-course transcriptomic information reveals the mechanisms of improved drought tolerance by drought priming in wheat[J]. >Journal of Integrative Agriculture, 2025, 24(8): 2902-2919.
[2] Shuran Li, Chunqing Ou, Fei Wang, Yanjie Zhang, Omayma Ismail, Yasser S. G. Abd Elaziz, Sherif Edris, , He Li, Shuling Jiang. Ppbbx24-del mutant positively regulates light-induced anthocyanin accumulation in the ‘Red Zaosu’ pear (Pyrus pyrifolia White Pear Group)[J]. >Journal of Integrative Agriculture, 2025, 24(7): 2619-2639.
[3] Congrui Sun, Runze Wang, Jiaming Li, Xiaolong Li, Bobo Song, David Edwards, Jun Wu. Pan-transcriptome analysis provides insights into resistance and fruit quality breeding of pear (Pyrus pyrifolia)[J]. >Journal of Integrative Agriculture, 2025, 24(5): 1813-1830.
[4] Xiaomei Tang, Yue Wang, Yuqing Guo, Luoluo Xie, Wei Song, Ziwen Xiao, Ruichang Yin, Zhe Ye, Xueqiu Sun, Wenming Wang, Lun Liu, Zhenfeng Ye, Zhenghui Gao, Bing Jia. Integrated transcriptomic and metabolomic analyses reveal a novel mechanism of resistance to Colletotrichum fructicola in pear[J]. >Journal of Integrative Agriculture, 2025, 24(10): 3851-3865.
[5] Meiyu Li, Liyun Mao, Shuang Song, Decang Kong, Ming Cao, Lijun Jiao, Wenhao Bo, Xiaoming Pang. Functional insights into the FW2.2-like gene family in Chinese jujube: Identification, characterization, and impact of ZjFWL10 variants on fruit size and plant height[J]. >Journal of Integrative Agriculture, 2025, 24(10): 3880-3894.
[6] Fuli Gao, Zidong Wang, Wankun Liu, Min Liu, Baoyi Wang, Yingjie Yang, Jiankun Song, Zhenhua Cui, Chenglin Liang, Dingli Li, Ran Wang, Jianlong Liu. Dehydrin PbDHN3 regulates ethylene synthesis and signal transduction to improve salt tolerance in pear[J]. >Journal of Integrative Agriculture, 2025, 24(10): 3838-3850.
[7] Lijiao Ge, Weihao Miao, Kuolin Duan, Tong Sun, Xinyan Fang, Zhiyong Guan, Jiafu Jiang, Sumei Chen, Weimin Fang, Fadi Chen, Shuang Zhao. Comparative transcriptome analysis identifies key regulators of nitrogen use efficiency in chrysanthemum[J]. >Journal of Integrative Agriculture, 2025, 24(1): 176-195.
[8] Tong Shen, Mengdi Ye, Yeping Xu, Bohan Ding, Hongtao Li, Li Zhang, Jun Wang, Yanli Tian, Baishi Hu, Youfu Zhao. Cytospora pyri promotes Erwinia amylovora virulence by providing metabolites and hyphae[J]. >Journal of Integrative Agriculture, 2024, 23(9): 3045-3054.
[9] Zhengyuan Xu, Lingzhen Ye, Qiufang Shen, Guoping Zhang. Advances in the study of waterlogging tolerance in plants[J]. >Journal of Integrative Agriculture, 2024, 23(9): 2877-2897.
[10] Guoling Guo, Haiyan Zhang, Weiyu Dong, Bo Xu, Youyu Wang, Qingchen Zhao, Lun Liu, Xiaomei Tang, Li Liu, Zhenfeng Ye, Wei Heng, Liwu Zhu, Bing Jia. Overexpression of PbrGA2ox1 enhances pear drought tolerance through the regulation of GA3-inhibited reactive oxygen species detoxification and abscisic acid signaling[J]. >Journal of Integrative Agriculture, 2024, 23(9): 2989-3011.
[11] Wanting Yu, Xinnan Zhang, Weiwei Yan, Xiaonan Sun, Yang Wang, Xiaohui Jia. Effects of 1-methylcyclopropene on skin greasiness and quality of ‘Yuluxiang’ pear during storage at 20°C[J]. >Journal of Integrative Agriculture, 2024, 23(7): 2476-2490.
[12] YANG Wei-bing, ZHANG Sheng-quan, HOU Qi-ling, GAO Jian-gang, WANG Han-Xia, CHEN Xian-Chao, LIAO Xiang-zheng, ZHANG Feng-ting, ZHAO Chang-ping, QIN Zhi-lie.

Transcriptomic and metabolomic analysis provides insights into lignin biosynthesis and accumulation and differences in lodging resistance in hybrid wheat [J]. >Journal of Integrative Agriculture, 2024, 23(4): 1105-1117.

[13] Xiaoyan Cui, Ke Yang, Weiyun Zhang, Liyang Zhang, Ding Li, Wei Wu, Yun Hu, Tingting Li, Xugang Luo. Dietary manganese supplementation inhibits abdominal fat deposition possibly by regulating gene expression and enzyme activity involved in lipid metabolism in the abdominal fat of broilers[J]. >Journal of Integrative Agriculture, 2024, 23(12): 4161-4171.
[14] Muhammad Mobeen Tahir, Li Fan, Zhimin Liu, Humayun Raza, Usman Aziz, Asad Shehzaib, Shaohuan Li, Yinnan He, Yicen Lu, Xiaoying Ren, Dong Zhang, Jiangping Mao. Physiological and molecular mechanisms of cytokinin involvement in nitrate-mediated adventitious root formation in apples[J]. >Journal of Integrative Agriculture, 2024, 23(12): 4046-4057.
[15] Atiqur RAHMAN, Md. Hasan Sofiur RAHMAN, Md. Shakil UDDIN, Naima SULTANA, Shirin AKHTER, Ujjal Kumar NATH, Shamsun Nahar BEGUM, Md. Mazadul ISLAM, Afroz NAZNIN, Md. Nurul AMIN, Sharif AHMED, Akbar HOSAIN. Advances in DNA methylation and its role in cytoplasmic male sterility in higher plants[J]. >Journal of Integrative Agriculture, 2024, 23(1): 1-19.
No Suggested Reading articles found!