Chinese Journal OF Rice Science >
Characterization and Gene Mapping of a No-pollen Genic Sterile Mutant whf41 in Rice
Received date: 2016-12-01
Revised date: 2017-02-06
Online published: 2017-05-10
【Objective】The anther cuticle and wax layer act as structural support and protective barriers for the development of pollen sac in rice. Phenotypic analysis and genetic mapping of the related genes can provide genetic resources and lay theoretical basis for further cloning and molecular mechanism analysis. 【Method】We identified a no-pollen male sterile mutant whf41 from the mutant progeny of 60Co-γ-treated indica cultivar Zhonghui 8015. Anthers of the wild-type and whf41 mutant at different development stages were fixed for transverse section analysis and mature anthers of which were randomly selected for scanning electron microscopy analysis. BC1F1, F1 lines and BC1F2, F2 populations derived from whf41/ Zhonghui 8015 and whf41/ 02428 were used for genetic analysis and fine-mapping of the mutation site. 【Result】 Comparing with wild type,the whf41 mutant exhibited thin and transparent milky anther without mature pollen grains. Transverse section analysis showed that the whf41 mutant displayed aborted pollen grains,none detectable exine and swollen tapetal layer without expected PCD (programmed cell death), finally resulting in fragments of tapetal layer and pollen grains in the chamber. Scanning electron microscopy analysis further revealed the mutant had smoother anther wall, fewer lipids both on the inner and outer sides and fragments of degragated pollens. Genetic analysis revealed that whf41 was controlled by a single recessive genic gene, which was fine-mapped to a 45.6-kb region that harbored nine Open Reading Frames(ORFs) on the short arm of chromosome 3 between markers XD-5 and XD-11. Sequence analysis revealed that the whf41 mutant carried a nucleotide substitution and three nucleotide deletion, which led a substitution of one amino acid (D to M)and a deletion of one amino acid (V) in the fourth exon of one Cytochrome P450 family gene, LOC_Os03g0725, which is named as OsWHF41 and probably responsible for the no-pollen male sterility phenotype. Furthermore, qPCR analysis showed that the expression level of CYP704B2 and seven anther lipid synthesis and transportation related regulators changed significantly in whf41. 【Conclusion】Based on all these results, we deduced that the OsWHF41 gene was a new allelic to CYP704B2 gene and this further confirmed the important role of CYP704B2 in anther lipid synthesis and pollen exine formation .
Key words： rice; whf41; recessive genic male sterility; gene mapping
Dandan XUAN, Lianping SUN, Peipei ZHANG, Yingxin ZHANG, Weixun WU, Zhengfu YANG, Xiaodeng ZHAN, Xihong SHEN, Liyong CAO, Shihua CHENG . Characterization and Gene Mapping of a No-pollen Genic Sterile Mutant whf41 in Rice[J]. Chinese Journal OF Rice Science, 2017 , 31(3) : 247 -256 . DOI: 10.16819/j.1001-7216.2017.6159 247
| [1] | 朱英国. 水稻雄性不育生物学. 武汉: 武汉大学出版社, 2000: 1-2. |
| [1] | Zhu Y G.Biology of Male Sterility in Rice. Wuhan: Wuhan University Press, 2000: 1-2. (in Chinese) |
| [2] | Raghavan V.Anther developmental biology in molecular embryology of flowering plants.Plant Physiol, 1997: 17-60. |
| [3] | Ma H.Molecular genetic analyses of microsporogenesis and microgametogenesis in flowering plants.Annu Rev Plant Boil, 2005, 56: 393-434. |
| [4] | Zhang D B, Wilson Z A.Stamen specification and anther development in rice. Chin Sci Bull, 2009, 54: 2342-2353. |
| [5] | Zhang D B, Luo X, Zhu L.Cytological analysis and genetic control of rice anther development.J Gen Genom, 2011, 38(9): 379-390. |
| [6] | Kinoshita T.Gene symbols and information on male sterility.Rice Genet Newsl, 1997, 14: 13-22. |
| [7] | Okamuro J K, Boer B G, Jofuku K D.Regulation of Arabidopsis flower development.Plant Cell, 1993, 5: 1183-1193. |
| [8] | 马西青, 方才臣, 邓联武, 万向元. 水稻隐性核雄性不育研究进展及育种应用探讨.中国水稻科学,2012, 25(5): 511-520. |
| [8] | Ma X Q, Fang C C, Deng L W, Wan X Y.Research progress and breeding application of recessive genic male sterility gene in rice.Chin J Rice Sci, 2012, 25(5): 511-520. (in Chinese with English abstract) |
| [9] | Nonomura K I, Miyoshi K, Eiguchi M, Suzuki T, Miyao A, Hirochika H, Kurata N.The MSP1 gene is necessary to restrict the number of cells entering into male and female sporogenesis and to initiate anther wall formation in rice.Plant Cell, 2003, 15(8): 1728-1739. |
| [10] | Nonomura K I, Nakano M, Fukuda T, Eiguchi M, Miyao A, Hirochika H, Kurata N.The novel gene HOMOLOGOUS PAIRING ABERRATION IN RICE MEIOSIS1 of rice encodes a putative coiled-coil protein required for homologous chromosome pairing in meiosis.Plant J, 2004, 16(4): 1008-1020. |
| [11] | Nonomura K I, Nakano M, Murata K, Miyoshi K, Eiguchi M, Miyao A, Hirochika H, Kurata N.An insertional mutation in the rice PAIR2 gene, the ortholog of Arabidopsis ASY1, results in a defect in homologous chromosome pairing during meiosis.Mol Gen Genom, 2004, 271(2): 121-129. |
| [12] | Yuan W Y, Li X W, Chang Y X, Wen R Y, Chen G X, Zhang Q F, Wu C.Mutation of the rice gene PAIR3 results in lack of bivalent formation in meiosis.Plant J, 2009, 59(2): 303-315. |
| [13] | Wang M, Wang K J, Tang D, Wei C X, Li M, Shen Y, Chi Z C, Gu M H, Cheng Z K.The central element protein ZEP1 of the synaptonemal complex regulates the number of crossovers during meiosis in rice.Plant Cell, 2010, 22: 417-430. |
| [14] | Nonomura K, Morohoshi A, Nakano M, Eiguchi M, Miyao A, Hirochika H, Kurata N.A germ cell-specific gene of the ARGONAUTE family is essential for the progression of premeiotic mitosis and meiosis during sporogenesis in rice.Plant Cell, 2007, 19(8): 2583-2594. |
| [15] | Wang C, Yue W, Ying Y, Wang S, Secco D, Liu Y, Whelan J, Tyerman S D, Shou H.Rice SPX-Major facility superfamily3, a vacuolar phosphate efflux transporter, is involved in maintaining phosphate homeostasis in rice.Plant Physiol, 2015, 169(4): 2822-2831. |
| [16] | He Y, Wang C, Higgins J D, Yu J, Zong J, Lu P, Zhang D, Liang W.MEIOTIC F-BOX is essential for male meiotic DNA double-strand break repair in rice.Plant Cell, 2016, 28(8): 1879-1893. |
| [17] | Li L, Li Y, Song S, Deng H, Li N, Fu X, Chen G, Yuan L.An anther development F-box (ADF) protein regulated by tapetum degeneration retardation (TDR) controls rice anther development.Planta, 2015, 241(1): 157-166. |
| [18] | Jung K H, Han M J, Lee Y S, Kim Y W, Hwang I, Kim M J, Kim Y K, Nahm B H, An G.Rice Undeveloped Tapetum1 is a major regulator of early tapetum development.Plant Cell, 2005, 17(10): 2705-2722. |
| [19] | Liu Z, Bao W, Liang W, Yin J, Zhang D.Identification of gamyb-4 and analysis of the regulatory role of GAMYB in rice anther development.J Integr Plant Biol, 2010, 52(7): 670-678. |
| [20] | Li H, Yuan Z, Vizcay-Barrena G, Yang C, Liang W, Zong J, Wilson Z A, Zhang D.PERSISTENT TAPETAL CELL1 encodes a PHD-Finger protein that is required for tapetal cell death and pollen development in rice.Plant Physiol, 2011, 156(2): 615-630. |
| [21] | Li X, Gao X, Wei Y, Deng L, Ouyang Y, Chen G, Li X, Zhang Q, Wu C.Rice APOPTOSIS INHIBITOR5 coupled with two DEAD-box adenosine 5′-triphosphate- dependent RNA helicases regulates tapetum degeneration.Plant Cell, 2011, 23(4): 1416-1434. |
| [22] | Niu N, Liang W, Yang X, Jin W, Wilson Z A, Hu J, Zhang D.EAT1 promotes tapetal cell death by regulating aspartic proteases during male reproductive development in rice. Nat Comm, 2013, 4: 1445. |
| [23] | Fu Z, Yu J, Cheng X, Zong X, Xu J, Chen M, Li Z, Zhang D, Liang W.The rice basic Helix-Loop-Helix transcription factor TDR INTERACTING PROTEIN2 is a central switch in early anther development.Plant Cell, 2014, 26(4): 1512-1524. |
| [24] | Niu B X, He F R, He M, Ren D, Chen L T, Liu Y G.The ATP-binding cassette transporter OsABCG15 is required for anther development and pollen fertility in rice.J Integr Plant Biol, 2013, 55(8): 710-720. |
| [25] | Zhao G, Shi J, Liang W, Xue F, Luo Q, Zhu L, Qu G, Chen M, Schreiber L, Zhang D.Two ATP binding cassette G transporters, rice ATP binding cassette G26 and ATP binding cassette G15, collaboratively regulate rice male reproduction.Plant Physiol, 2015, 169(3): 2064-2079. |
| [26] | Zhang D, Liang W, Yin C, Zong J, Gu F, Zhang D.OsC6, encoding a lipid transfer protein, is required for postmeiotic anther development in rice.Plant Physiol, 2010, 154(1): 149-162. |
| [27] | Jung K H, Han M J, Lee D Y, Lee Y S, Schreiber L, Franke R, Faust A, Yephremov A, Saedler H, Kim Y W, Hwang I, An G.Wax-Deficient Anther1 is involved in cuticle and wax production in rice anther walls and is required for pollen development.Plant Cell, 2006, 18(11): 3015-3032. |
| [28] | Shi J, Tan H, Yu X H, Liu Y, Liang W, Ranathunge K, Franke R B, Schreiber L, Wang Y, Kai G, Shanklin J, Ma H, Zhang D.Defective Pollen Wall is required for anther and microspore development in rice and encodes a fatty acyl carrier protein reductase.Plant Cell, 2011, 23(6): 2225-2246. |
| [29] | Xu D W, Shi J X, Rautengarten C, Yang L, Qian X L, Uzair M, Zhu L, Luo Q, An G, Waßmann F, Schreiber L, Heazlewood J L, Scheller H V, Hu J P, Zhang D B, Liang W Q.Defective Pollen Wall 2 (DPW2) encodes an acyl transferase required for rice pollen development.Plant Physiol, 2017, 173(1): 240-255. |
| [30] | Li H, Pinot F, Sauveplane V, Werck-Reichhart D, Diehl P, Schreiber L, Franke R, Zhang P, Chen L, Gao Y, Liang W, Zhang D.Cytochrome P450 family member CYP704B2 catalyzes the ω -hydroxylation of fatty acids and is required for anther cutin biosynthesis and pollen exine formation in rice. Plant Cell, 2010, 22(1): 173-190. |
| [31] | Yang X, Wu D, Shi J, He Y, Pinot F, Grausem B, Yin C, Zhu L, Chen M, Luo Z, Liang W, Zhang D.Rice CYP703A3, a cytochrome P450 hydroxylase, is essential for development of anther cuticle and pollen exine.J Integr Plant Biol, 2014, 56(10): 979-994. |
| [32] | Zhu Q H, Ramm K, Shivakkumar R, Dennis E S, Upadhyaya N M.The ANTHER INDEHISCENCE1 gene encoding a single MYB domain protein is involved in anther development in rice.Plant Physiol, 2004, 135(3): 1514-1525. |
| [33] | Kent B, Erin C, Nina D, David H, Young J K, Zhong Cathy X Y. Plant genomic DNA flanking SPT event and methods for identifying SPT event. United States Patent, US008257930B2, 20120904. |
| [34] | Chang Z Y, Chen Z F, Wang N, Xie G, Lu J W, Yan W, Zhou J L, Tang X Y, Deng X W.Construction of a male sterility system for hybrid rice breeding and seed production using a nuclear male sterility gene.Proc Natl Acad Sci USA, 2016, 113(49): 14145. |
| [35] | Sun L P, Zhang Y X, Zhang P P, Yang Z F, Zhan X D, Shen X H, Zhang Z H, Hu X, Xuan D D, Wu W X, Li Z H, Cao L Y, Cheng S H.K-Domain splicing factor OsMADS1 regulates open hull male sterility in rice.Rice Sci, 2015, 22(5): 207-216. |
| [36] | Orjuela J, Garavito A, Bouniol M, Arbelaez J D, Moreno L, Kimball J, Wilson G, Rami J F, Tohme J, McCouch S R, Lorieux M. A universal core genetic map for rice.Theor Appl Genet, 2010, 120: 563-572 |
| [37] | Wu D H, Wu H P, Wang C S, Tseng H Y, Hwu K K.Genome-wide InDel marker system for application in rice breeding and mapping studies.Euphytica, 2013, 192: 131-143 |
| [38] | Yamanaka S, Suzuki E, Tanaka M, Takeda Y, Watanabe J A, Watanabe K N.Assessment of cytochrome P450 sequences offers a useful tool for determining genetic diversity in higher plant species.Theor Appl Genet, 2003, 108(1): 1-9. |
| [39] | Tang W, Wu T, Ye J, Sun J, Jiang Y, Yu J, Tang J, Chen G, Wang C, Wan J.SNP-based analysis of genetic diversity reveals important alleles associated with seed size in rice.BMC Plant Biol, 2016, 16(1): 93. |
/
| 〈 |
|
〉 |