Techniques of cell type-specific transcriptome analysis and applications in researches of sexual plant reproduction
Tian-Xiang HU, Miao YU, Jie ZHAO
Techniques of cell type-specific transcriptome analysis and applications in researches of sexual plant reproduction
In higher plants, specific cell differentiation and fate decision are controlled by differential gene expression. Cell type-specific transcriptome analysis has become an important tool for investigating cell regulatory mechanisms. In recent years, many different techniques have been developed for the isolation of specific cells and the subsequent transcriptome analysis, and considerable data are available regarding the transcriptional profiles of some specific cells. These cell type-specific transcriptome analyses hold significant promise for elucidating the gene expression linked to cellular identities and functions, and are extraordinarily important for research in functional genomics and systems biology aimed toward basic understanding of molecular networks and pathway interactions. Moreover, to reveal the critical mechanisms about sexual plant reproduction, the gamete and embryo cells have long been treated as good subjects for cell-specific transcriptome analysis, and there has been important progress in recent decades. In this review, we summarize current technologies in cell type-specific transcriptome analysis and review the applications of these technologies in research into the mechanisms of sexual reproduction in higher plants.
specific cell / transcriptome / plant reproduction
[1] |
Aharoni A, Vorst O (2002). DNA microarrays for functional plant genomics. Plant Mol Biol, 48(1-2): 99-118
CrossRef
Pubmed
Google scholar
|
[2] |
Ansorge W J (2009). Next-generation DNA sequencing techniques. New Biotechnol, 25(4): 195-203
CrossRef
Pubmed
Google scholar
|
[3] |
Asano T, Masumura T, Kusano H, Kikuchi S, Kurita A, Shimada H, Kadowaki K (2002). Construction of a specialized cDNA library from plant cells isolated by laser capture microdissection: toward comprehensive analysis of the genes expressed in the rice phloem. Plant J, 32(3): 401-408
CrossRef
Pubmed
Google scholar
|
[4] |
Baginsky S, Gruissem W (2006). Arabidopsis thaliana proteomics: from proteome to genome. J Exp Bot, 57(7): 1485-1491
CrossRef
Pubmed
Google scholar
|
[5] |
Becker J D, Boavida L C, Carneiro J, Haury M, Feijó J A (2003). Transcriptional profiling of Arabidopsis tissues reveals the unique characteristics of the pollen transcriptome. Plant Physiol, 133(2): 713-725
CrossRef
Pubmed
Google scholar
|
[6] |
Belyavsky A, Vinogradova T, Rajewsky K (1989). PCR-based cDNA library construction: general cDNA libraries at the level of a few cells. Nucleic Acids Res, 17(8): 2919-2932
CrossRef
Pubmed
Google scholar
|
[7] |
Birnbaum K, Shasha D E, Wang J Y, Jung J W, Lambert G M, Galbraith D W, Benfey P N (2003). A gene expression map of the Arabidopsis root. Science, 302(5652): 1956-1960
CrossRef
Pubmed
Google scholar
|
[8] |
Borges F, Gomes G, Gardner R, Moreno N, McCormick S, Feijó J A, Becker J D (2008). Comparative transcriptomics of Arabidopsis sperm cells. Plant Physiol, 148(2): 1168-1181
CrossRef
Pubmed
Google scholar
|
[9] |
Brady S M, Orlando D A, Lee J Y, Wang J Y, Koch J, Dinneny J R, Mace D, Ohler U, Benfey P N (2007). A high-resolution root spatiotemporal map reveals dominant expression patterns. Science, 318(5851): 801-806
CrossRef
Pubmed
Google scholar
|
[10] |
Brandt S, Kehr J, Walz C, Imlau A, Willmitzer L, Fisahn J (1999). Technical advance: A rapid method for detection of plant gene transcripts from single epidermal, mesophyll and companion cells of intact leaves. Plant J, 20(2): 245-250
CrossRef
Pubmed
Google scholar
|
[11] |
Brandt S, Kloska S, Altmann T, Kehr J (2002). Using array hybridization to monitor gene expression at the single cell level. J Exp Bot, 53(379): 2315-2323
CrossRef
Pubmed
Google scholar
|
[12] |
Cao Y, Reece A, Russell S D (1996). Isolation of viable sperm cells from tobacco (Nicotiana tabacum). Zygote, 4(2): 81-84
CrossRef
Pubmed
Google scholar
|
[13] |
Casson S, Spencer M, Walker K, Lindsey K (2005). Laser capture microdissection for the analysis of gene expression during embryogenesis of Arabidopsis. Plant J, 42(1): 111-123
CrossRef
Pubmed
Google scholar
|
[14] |
Chen S H, Yang Y H, Liao J P, Kuang A X, Tian H Q (2008). Isolation of egg cells and zygotes of Torenia fournieri L. and determination of their surface charge. Zygote, 16(2): 179-186
CrossRef
Pubmed
Google scholar
|
[15] |
Diatchenko L, Lau Y F, Campbell A P, Chenchik A, Moqadam F, Huang B, Lukyanov S, Lukyanov K, Gurskaya N, Sverdlov E D, Siebert P D (1996). Suppression subtractive hybridization: a method for generating differentially regulated or tissue-specific cDNA probes and libraries. Proc Natl Acad Sci USA, 93(12): 6025-6030
CrossRef
Pubmed
Google scholar
|
[16] |
Dresselhaus T, Lörz H, Kranz E (1994). Representative cDNA libraries from few plant cells. Plant J, 5(4): 605-610
CrossRef
Pubmed
Google scholar
|
[17] |
Elling A A, Deng X W (2009). Next-generation sequencing reveals complex relationships between the epigenome and transcriptome in maize. Plant Signal Behav, 4(8): 760-762
CrossRef
Pubmed
Google scholar
|
[18] |
Emmert-Buck M R, Bonner R F, Smith P D, Chuaqui R F, Zhuang Z, Goldstein S R, Weiss R A, Liotta L A (1996). Laser capture microdissection. Science, 274(5289): 998-1001
CrossRef
Pubmed
Google scholar
|
[19] |
Engel M L, Chaboud A, Dumas C, McCormick S (2003). Sperm cells of Zea mays have a complex complement of mRNAs. Plant J, 34(5): 697-707
CrossRef
Pubmed
Google scholar
|
[20] |
Fu C M, Sun M X, Zhou C, Yang H Y (1996). Isolation of fertilized embryo sacs and zygotes and triggering of zygote division in vitro in Nicotiana tabacum. Acta Bot Sin, 38: 262-267
|
[21] |
Ge X, Chen W, Song S, Wang W, Hu S, Yu J (2008). Transcriptomic profiling of mature embryo from an elite super-hybrid rice LYP9 and its parental lines. BMC Plant Biol, 8(1): 114
CrossRef
Pubmed
Google scholar
|
[22] |
Givan A L (2001). Flow Cytometry First Principles.New York: Wiley-Liss,. 273
|
[23] |
Goldberg R B, Hoschek G, Ditta G S, Breidenbach R W (1981). Developmental regulation of cloned superabundant embryo mRNAs in soybean. Dev Biol, 83(2): 218-231
CrossRef
Pubmed
Google scholar
|
[24] |
Gou X, Yuan T, Wei X, Russell S D (2009). Gene expression in the dimorphic sperm cells of Plumbago zeylanica: transcript profiling, diversity, and relationship to cell type. Plant J, 60(1): 33-47
CrossRef
Pubmed
Google scholar
|
[25] |
Gou X P, Xu Y, Tang L, Yan F, Chen F (2001). Representative cDNA library from isolated rice sperm cells. Acta Bot Sin, 43: 1093-1096
|
[26] |
Haerizadeh F, Wong C E, Bhalla P L, Gresshoff P M, Singh M B (2009). Genomic expression profiling of mature soybean (Glycine max) pollen. BMC Plant Biol, 9(1): 25
CrossRef
Pubmed
Google scholar
|
[27] |
Herzenberg L A, De Rosa S C (2000). Monoclonal antibodies and the FACS: complementary tools for immunobiology and medicine. Immunol Today, 21(8): 383-390
CrossRef
Pubmed
Google scholar
|
[28] |
Hobo T, Suwabe K, Aya K, Suzuki G, Yano K, Ishimizu T, Fujita M, Kikuchi S, Hamada K, Miyano M, Fujioka T, Kaneko F, Kazama T, Mizuta Y, Takahashi H, Shiono K, Nakazono M, Tsutsumi N, Nagamura Y, Kurata N, Watanabe M, Matsuoka M (2008). Various spatiotemporal expression profiles of anther-expressed genes in rice. Plant Cell Physiol, 49(10): 1417-1428
CrossRef
Pubmed
Google scholar
|
[29] |
Holm P B, Knudsen S, Mouritzen P, Negri D, Olsen F L, Roue C (1994). Regeneration of fertile barley plants from mechanically isolated protoplasts of the fertilized egg cell. Plant Cell, 6(4): 531-543
CrossRef
Pubmed
Google scholar
|
[30] |
Honys D, Twell D (2003). Comparative analysis of the Arabidopsis pollen transcriptome. Plant Physiol, 132(2): 640-652
CrossRef
Pubmed
Google scholar
|
[31] |
Honys D, Twell D (2004). Transcriptome analysis of haploid male gametophyte development in Arabidopsis. Genome Biol, 5(11): R85
CrossRef
Pubmed
Google scholar
|
[32] |
Hu S Y, Li L G, Zhu C (1985). Isolation of viable embryo sacs and their protoplasts of Nicotiana tabacum. Acta Bot Sin, 27: 337-344
|
[33] |
Hu S Y, Yang H Y (2002). Biology of Angiosperm Fertilization.Beijing: Science Press, 250
|
[34] |
Hu T X, Yu M, Zhao J (2010). Comparative transcriptional profiling analysis of the two daughter cells from tobacco zygote reveals the transcriptome differences in the apical and basal cells. BMC Plant Biol, 10(1): 167
CrossRef
Pubmed
Google scholar
|
[35] |
Huang B Q, Russell S D (1989). Isolation of fixed and viable eggs, central cells, and embryo sacs from ovules of Plumbago zeylanica. Plant Physiol, 90(1): 9-12
CrossRef
Pubmed
Google scholar
|
[36] |
Ito Y, Chujo A, Eiguchi M, Kurata N (2004). Radial axis differentiation in a globular embryo is marked by HAZ1, a PHD-finger homeobox gene of rice. Gene, 331: 9-15
CrossRef
Pubmed
Google scholar
|
[37] |
Johnston A J, Meier P, Gheyselinck J, Wuest S E, Federer M, Schlagenhauf E, Becker J D, Grossniklaus U (2007). Genetic subtraction profiling identifies genes essential for Arabidopsis reproduction and reveals interaction between the female gametophyte and the maternal sporophyte. Genome Biol, 8(10): R204
CrossRef
Pubmed
Google scholar
|
[38] |
Jones-Rhoades M W, Borevitz J O, Preuss D (2007). Genome-wide expression profiling of the Arabidopsis female gametophyte identifies families of small, secreted proteins. PLoS Genet, 3(10): 1848-1861
CrossRef
Pubmed
Google scholar
|
[39] |
Karrer E E, Lincoln J E, Hogenhout S, Bennett A B, Bostock R M, Martineau B, Lucas W J, Gilchrist D G, Alexander D (1995). In situ isolation of mRNA from individual plant cells: creation of cell-specific cDNA libraries. Proc Natl Acad Sci U S A, 92(9): 3814-3818
CrossRef
Pubmed
Google scholar
|
[40] |
Katoh N, Lörz H, Kranz E (1997). Isolation of viable egg cells of rape (Brassica napus L.). Zygote, 5(1): 31-33
CrossRef
Pubmed
Google scholar
|
[41] |
Kerk N M, Ceserani T, Tausta S L, Sussex I M, Nelson T M (2003). Laser capture microdissection of cells from plant tissues. Plant Physiol, 132(1): 27-35
CrossRef
Pubmed
Google scholar
|
[42] |
Kranz E, Bautor J, Lörz H (1991). In vitro fertilization of single, isolated gametes of maize mediated by electrofusion. Sex Plant Reprod, 4: 12-16
CrossRef
Google scholar
|
[43] |
Kumlehn J, Lörz H, Kranz E (1998). Differentiation of isolated wheat zygotes into embryos and normal plants. Planta, 205(3): 327-333
CrossRef
Google scholar
|
[44] |
Kumlehn K, Czihal A, Matzk L (2001). Parthenogenetic egg cells of wheat: cellular and molecular studies. Sex Plant Reprod, 14(4): 239-243
CrossRef
Google scholar
|
[45] |
Le B H, Cheng C, Bui A Q, Wagmaister J A, Henry K F, Pelletier J, Kwong L, Belmonte M, Kirkbride R, Horvath S, Drews G N, Fischer R L, Okamuro J K, Harada J J, Goldberg R B (2010). Global analysis of gene activity during Arabidopsis seed development and identification of seed-specific transcription factors. Proc Natl Acad Sci USA, 107(18): 8063-8070
CrossRef
Pubmed
Google scholar
|
[46] |
Le B H, Wagmaister J A, Kawashima T, Bui A Q, Harada J J, Goldberg R B (2007). Using genomics to study legume seed development. Plant Physiol, 144(2): 562-574
CrossRef
Pubmed
Google scholar
|
[47] |
Lê Q, Gutièrrez-Marcos J F, Costa L M, Meyer S, Dickinson H G, Lörz H, Kranz E, Scholten S (2005). Construction and screening of subtracted cDNA libraries from limited populations of plant cells: a comparative analysis of gene expression between maize egg cells and central cells. Plant J, 44(1): 167-178
CrossRef
Pubmed
Google scholar
|
[48] |
Lee J M, Williams M E, Tingey S V, Rafalski J A (2002). DNA array profiling of gene expression changes during maize embryo development. Funct Integr Genomics, 2(1-2): 13-27
CrossRef
Pubmed
Google scholar
|
[49] |
Loader N M, Woolner E M, Hellyer A, Slabas A R, Safford R (1993). Isolation and characterization of two Brassica napus embryo acyl-ACP thioesterase cDNA clones. Plant Mol Biol, 23(4): 769-778
CrossRef
Pubmed
Google scholar
|
[50] |
Márton M L, Cordts S, Broadhvest J, Dresselhaus T (2005). Micropylar pollen tube guidance by egg apparatus 1 of maize. Science, 307(5709): 573-576
CrossRef
Pubmed
Google scholar
|
[51] |
Nakazono M, Qiu F, Borsuk L A, Schnable P S (2003). Laser-capture microdissection, a tool for the global analysis of gene expression in specific plant cell types: identification of genes expressed differentially in epidermal cells or vascular tissues of maize. Plant Cell, 15(3): 583-596
CrossRef
Pubmed
Google scholar
|
[52] |
Nelson T, Tausta S L, Gandotra N, Liu T (2006). Laser microdissection of plant tissue: what you see is what you get. Annu Rev Plant Biol, 57(1): 181-201
CrossRef
Pubmed
Google scholar
|
[53] |
Ning J, Peng X B, Qu L H, Xin H P, Yan T T, Sun M (2006). Differential gene expression in egg cells and zygotes suggests that the transcriptome is restructed before the first zygotic division in tobacco. FEBS Lett, 580(7): 1747-1752
CrossRef
Pubmed
Google scholar
|
[54] |
Okada T, Bhalla P L, Singh M B (2006). Expressed sequence tag analysis of Lilium longiflorum generative cells. Plant Cell Physiol, 47(6): 698-705
CrossRef
Pubmed
Google scholar
|
[55] |
Okada T, Singh M B, Bhalla P L (2007). Transcriptome profiling of Lilium longiflorum generative cells by cDNA microarray. Plant Cell Rep, 26(7): 1045-1052
CrossRef
Pubmed
Google scholar
|
[56] |
Okamoto T, Higuchi K, Shinkawa T, Isobe T, Lörz H, Koshiba T, Kranz E (2004). Identification of major proteins in maize egg cells. Plant Cell Physiol, 45(10): 1406-1412
CrossRef
Pubmed
Google scholar
|
[57] |
Okamoto T, Scholten S, Lörz H, Kranz E (2005). Identification of genes that are up- or down-regulated in the apical or basal cell of maize two-celled embryos and monitoring their expression during zygote development by a cell manipulation- and PCR-based approach. Plant Cell Physiol, 46(2): 332-338
CrossRef
Pubmed
Google scholar
|
[58] |
Okuda S, Tsutsui H, Shiina K, Sprunck S, Takeuchi H, Yui R, Kasahara R D, Hamamura Y, Mizukami A, Susaki D, Kawano N, Sakakibara T, Namiki S, Itoh K, Otsuka K, Matsuzaki M, Nozaki H, Kuroiwa T, Nakano A, Kanaoka M M, Dresselhaus T, Sasaki N, Higashiyama T (2009). Defensin-like polypeptide LUREs are pollen tube attractants secreted from synergid cells. Nature, 458(7236): 357-361
CrossRef
Pubmed
Google scholar
|
[59] |
Pina C, Pinto F, Feijó J A, Becker J D (2005). Gene family analysis of the Arabidopsis pollen transcriptome reveals biological implications for cell growth, division control, and gene expression regulation. Plant Physiol, 138(2): 744-756
CrossRef
Pubmed
Google scholar
|
[60] |
Ranford J C, Bryce J H, Morris P C (2002). PM19, a barley (Hordeum vulgare L.) gene encoding a putative plasma membrane protein, is expressed during embryo development and dormancy. J Exp Bot, 53(366): 147-148
CrossRef
Pubmed
Google scholar
|
[61] |
Russell S D (1986). Isolation of sperm cells from the pollen of Plumbago zeylanica. Plant Physiol, 81(1): 317-319
CrossRef
Pubmed
Google scholar
|
[62] |
Russell S D (1992). Double fertilization. Int Rev Cytol, 40: 357-390
CrossRef
Google scholar
|
[63] |
Sanger F (1988). Sequences, sequences, and sequences. Annu Rev Biochem, 57(1): 1-28
CrossRef
Pubmed
Google scholar
|
[64] |
Shendure J, Ji H (2008). Next-generation DNA sequencing. Nat Biotechnol, 26(10): 1135-1145
CrossRef
Pubmed
Google scholar
|
[65] |
Simone N L, Bonner R F, Gillespie J W, Emmert-Buck M R, Liotta L A (1998). Laser-capture microdissection: opening the microscopic frontier to molecular analysis. Trends Genet, 14(7): 272-276
CrossRef
Pubmed
Google scholar
|
[66] |
Spencer M W, Casson S A, Lindsey K (2007). Transcriptional profiling of the Arabidopsis embryo. Plant Physiol, 143(2): 924-940
CrossRef
Pubmed
Google scholar
|
[67] |
Sprunck S, Baumann U, Edwards K, Langridge P, Dresselhaus T (2005). The transcript composition of egg cells changes significantly following fertilization in wheat (Triticum aestivum L.). Plant J, 41(5): 660-672
CrossRef
Pubmed
Google scholar
|
[68] |
Steffen J G, Kang I H, Macfarlane J, Drews G N (2007). Identification of genes expressed in the Arabidopsis female gametophyte. Plant J, 51(2): 281-292
CrossRef
Pubmed
Google scholar
|
[69] |
Stinson J R, Eisenberg A J, Willing R P, Pe M E, Hanson D D, Mascarenhas J P (1987). Genes expressed in the male gametophyte of flowering plants and their isolation. Plant Physiol, 83(2): 442-447
CrossRef
Pubmed
Google scholar
|
[70] |
Tanaka I (1988). Isolation of generative cells and their protoplasts from pollen of Lilium longiflorum. Protoplasma, 142(1): 68-73
CrossRef
Google scholar
|
[71] |
Tang F, Barbacioru C, Wang Y, Nordman E, Lee C, Xu N, Wang X, Bodeau J, Tuch B B, Siddiqui A, Lao K, Surani M A (2009). mRNA-Seq whole-transcriptome analysis of a single cell. Nat Methods, 6(5): 377-382
CrossRef
Pubmed
Google scholar
|
[72] |
Theerakulpisut P, Xu H, Singh M B, Pettitt J M, Knox R B (1991). Isolation and developmental expression of Bcp1, an anther-specific cDNA clone in Brassica campestris. Plant Cell, 3(10): 1073-1084
Pubmed
|
[73] |
Twell D, Wing R, Yamaguchi J, McCormick S (1989). Isolation and expression of an anther-specific gene from tomato. Mol Gen Genet, 217(2-3): 240-245
CrossRef
Pubmed
Google scholar
|
[74] |
Wall P K, Leebens-Mack J, Chanderbali A S, Barakat A, Wolcott E, Liang H, Landherr L, Tomsho L P, Hu Y, Carlson J E, Ma H, Schuster S C, Soltis D E, Soltis P S, Altman N, dePamphilis C W (2009). Comparison of next generation sequencing technologies for transcriptome characterization. BMC Genomics, 10(1): 347
CrossRef
Pubmed
Google scholar
|
[75] |
Wang D, Zhang C, Hearn D J, Kang I H, Punwani J A, Skaggs M I, Drews G N, Schumaker K S, Yadegari R (2010). Identification of transcription-factor genes expressed in the Arabidopsis female gametophyte. BMC Plant Biol, 10(1): 110
CrossRef
Pubmed
Google scholar
|
[76] |
Wang D Y, Zhang Q, Liu Y, Lin Z F, Zhang S X, Sun M X, Sodmergen (2010). The levels of male gametic mitochondrial DNA are highly regulated in angiosperms with regard to mitochondrial inheritance. Plant Cell, 22(7): 2402-2416
CrossRef
Pubmed
Google scholar
|
[77] |
Wei L Q, Xu W Y, Deng Z Y, Su Z, Xue Y, Wang T (2010). Genome-scale analysis and comparison of gene expression profiles in developing and germinated pollen in Oryza sativa. BMC Genomics, 11(1): 338
CrossRef
Pubmed
Google scholar
|
[78] |
Weterings K, Apuya N R, Bi Y, Fischer R L, Harada J J, Goldberg R B (2001). Regional localization of suspensor mRNAs during early embryo development. Plant Cell, 13(11): 2409-2425
Pubmed
|
[79] |
Whittle C A, Malik M R, Li R, Krochko J E (2010). Comparative transcript analyses of the ovule, microspore, and mature pollen in Brassica napus. Plant Mol Biol, 72(3): 279-299
CrossRef
Pubmed
Google scholar
|
[80] |
Wuest S E, Vijverberg K, Schmidt A, Weiss M, Gheyselinck J, Lohr M, Wellmer F, Rahnenführer J, von Mering C, Grossniklaus U (2010). Arabidopsis female gametophyte gene expression map reveals similarities between plant and animal gametes. Curr Biol, 20(6): 506-512
CrossRef
Pubmed
Google scholar
|
[81] |
Xin H P, Peng X B, Ning J, Yan T T, Ma L G, Sun M X (2010). Expressed sequence-tag analysis of tobacco sperm cells reveals a unique transcriptional profile and selective persistence of paternal transcripts after fertilization. Sex Plant Reprod,
CrossRef
Pubmed
Google scholar
|
[82] |
Xu H, Swoboda I, Bhalla P L, Sijbers A M, Zhao C, Ong E K, Hoeijmakers J H, Singh M B (1998). Plant homologue of human excision repair gene ERCC1 points to conservation of DNA repair mechanisms. Plant J, 13(6): 823-829
CrossRef
Pubmed
Google scholar
|
[83] |
Xu H, Swoboda I, Bhalla P L, Singh M B (1999). Male gametic cell-specific gene expression in flowering plants. Proc Natl Acad Sci USA, 96(5): 2554-2558
CrossRef
Pubmed
Google scholar
|
[84] |
Xu H, Weterings K, Vriezen W, Feron R, Xue Y, Derksen J, Mariani C (2002). Isolation and characterization of male-germ-cell transcripts in Nicotiana tabacum. Sex Plant Reprod, 14(6): 339-346
CrossRef
Google scholar
|
[85] |
Yang H, Kaur N, Kiriakopolos S, McCormick S (2006). EST generation and analyses towards identifying female gametophyte-specific genes in Zea mays L. Planta, 224(5): 1004-1014
CrossRef
Pubmed
Google scholar
|
[86] |
Yu H J, Hogan P, Sundaresan V (2005). Analysis of the female gametophyte transcriptome of Arabidopsis by comparative expression profiling. Plant Physiol, 139(4): 1853-1869
CrossRef
Pubmed
Google scholar
|
[87] |
Zhang G, Guo G, Hu X, Zhang Y, Li Q, Li R, Zhuang R, Lu Z, He Z, Fang X, Chen L, Tian W, Tao Y, Kristiansen K, Zhang X, Li S, Yang H, Wang J, Wang J (2010). Deep RNA sequencing at single base-pair resolution reveals high complexity of the rice transcriptome. Genome Res, 20(5): 646-654
CrossRef
Pubmed
Google scholar
|
[88] |
Zhang Z, Xu H, Singh M B, Russell S D (1998). Isolation and collection of two populations of viable sperm cells from the pollen of Plumbago zeylanica. Zygote, 6(4): 295-298
CrossRef
Pubmed
Google scholar
|
[89] |
Zhao J, Zhou C, Yang H Y (2000). Isolation and in vitro culture of zygotes and central cells of Oryza sativa L. Plant Cell Rep, 19(3): 321-326
CrossRef
Google scholar
|
/
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