1.     桃果实、花、根、叶：Isolation, characterisation and phylogenetic analysis of resistance gene    analogues in a wild species of peach (Prunus kansuensis).Canadian Journal of Plant Science,    2011, 91(6): 961-970

2.     樱桃花、叶、颚等各部位：Over-expression of the PaAP1 gene from sweet cherry (Prunus avium    L.)    causes early floweri.Journal of Plant Physiology, 2012,Available online 1 December    2012

3.     洋葱根、茎、蕾、叶、雌雄蕊等各部位：Cloning and Expression Analysis of A Putative B Class MADS-box    Gene of AcPI in Onion. Scientia Agricultura Sinica, 2012, 45(23):4759-4769

4.     芜菁：Isolation and Functional Characterisation of the Genes Encoding Δ8-Sphingolipid       Desaturase from Brassica rapa. Journal of Genetics and Genomics Volume 39, Issue 1, January    2012,    Pages 47–59

5.     芜  菁 1 ：EXPRESSION, DIVERGENCE AND EVOLUTION OF THE CALEOSIN GENE FAMILY IN BRASSICA RAPA. Arch. Biol. Sci., Belgrade, 65 (3), 863-876, 2013 DOI:10.2298/ABS1303863H

6.     番茄叶：Effect of Low Temperature Stress on the Expression of ProDH Gene and the Activities of the Proline Dehydrogenase in Leaves of Tomato Seedling. Chinese Agricultural Science  Bulletin 2012,28(10):132-135

7.     栀子叶：Isolation of High Quality Total RNA from Gardenia jasminoides Eills.Chinese Agricultural    Science Bulletin.2012, 28(27):194-198

8.     油桐果实：Cui Qinqin, Han Xiaojiao, Chen Yicun, Zhan Zhiyong, Lin Liyuan, Wang Yangdong.     Isolation and Expression Characteristics of Biotin Carboxyl Carrier Protein Coding Gene（VfBCCP)  from Vernicia fordii.SCIENTIA SILVAE SINICAE. 2012, 48(8): Available online August

9.     油桐果实1：Selection of Reliable Reference Genes for Gene Expression Studies Using Real-Time    PCR in Tung Tree during Seed Development. PLoS ONE, 2012, 7(8): e43084

10. 紫菜：Molecular cloning and expression analysis of ribosomal protein S7 gene from Porphyra    haitanensis. JOURNAL OF FISHERIES OF CHINA, 2011, 35（12）：1814-1821

11. 石斛：Molecular characterization of a mitogen-activated protein kinase gene DoMPK1 in Dendrobium officinale. Acta Pharmaceutica Sinica, 2012, 47 (12): 1703-1709

12. 石斛1：ESTs Analysis Reveals Putative Genes Involved in Symbiotic Seed Germination in Dendrobium officinale. Symbiotic Germination Genes in D. officinale. August 2013 | Volume 8 | Issue 8 | e72705

13. 大豆：RNA-seq Analysis Reveals Ethylene-Mediated Reproductive Organ Development and Abscission in Soybean(Glycine max L. Merr.). Plant Mol Biol Rep, 2012, published online: 4 Dec, 2012

14. 大豆1：Construction of ethylene regulatory network based on the phytohormones related gene transcriptome profiling and prediction of transcription factor activities in soybean. Acta Physiol Plant, 2012, published online: 12 Dec, 2012

15. 红花玉兰：Expression Analysis of MAwuAG in Different Organs and Developmental Stages of Magnolia wufengensis. Chinese Bulletin of Botany, 2013, 48 (2): 1–5

16. 毛桃：Cloning and Phylogeny Analysis of PpAP2 Floral Homologous Genes in Peach. Chinese Agricultural Science Bulletin, 2013, 29(7): 99-104

17. 五倍子：Cloning and characterisation of a phenylalanine ammonia-lyase gene from Rhus chinensis. Plant Cell Rep, 2013, published online：15 March, 2013

18. :五倍子1：Cloning, characterization and expression of chalcone synthase from medicinal plant Rhus chinensis.J. Plant Biochem. Biotechnol. DOI 10.1007/s13562-013-0231-9

19. 青杄 ：cDNA Cloning and Bioinformatic Analysis of the sPPa1 Gene form Picea wilsonii. Plant Science Journal, 2012, 30(40): 394-401

20. 青杄 1：cDNA Cloning and Bioinformatic Analysis of PsbO Gene from Picea wilsonii.Life Science Research, 2012, 16(3): 201-206

21. 青杄 2：Cloning and Tissue Expression Analysis of PwPSAF in Picea wilsonii. SCIENTIA SILVAE SINICAE. Vol. 49，No. 10, Oct. 2013.

22. 洋葱：Molecular Cloning and Transcriptional Analysis of the Putative AGAMOUS Homolog AcAG in Onion (Allium cepa. Plant Mol Biol Rep, DOI 10.1007/s11105-013-0607-y

23. 木瓜：XsFAD2 gene encodes the enzyme responsible for the high linoleic acid content in oil accumulated in Xanthoceras sorbifolia seeds. JOURNAL ARTICLE. 2013-6-17.

24. 木瓜1：Two novel diacylglycerol acyltransferase genes from Xanthoceras 2 sorbifolia are responsible for its seed oil content. GENE-38688; No. of pages: 9; 4C:

25. 柑橘：Efficient auto-excision of a selectable marker gene from transgenic citrus by combining the Cre/loxP system and ipt selection. Plant Cell Rep, DOI 10.1007/s00299-013-1470-x

26. 柑橘1：Expression Analysis of Three Phloem-specific Promoters in Transgenic Poncirus trifoliata. Acta Horticulturae Sinica. 2014, 41(1): 1–8.

27. 柑橘2： Activation of three pathogen-inducible promoters in transgenic citrus (Citrus sinensis Osbeck) after Xanthomonas axonopodis pv. citri infection and wounding. Plant Cell Tiss Organ Cult. DOI 10.1007/s11240-013-0423-y.

28. 茶梅花瓣：Comparison and Analysis of Methods of Extracting Total RNA from Petals of Camellia sasanqua. Chinese Agricultural Science Bulletin.2013,29(28):129-133.

29. 栀子：Isolation of High Quality Total RNA fromGardenia jasminoides Eills. Chinese Agricultural Science Bulletin. 2012, 28(27):194-198

30. 丹参：Genome-wide analysis and molecular dissection of the SPL gene family in Salvia miltiorrhiza. 2014 Jan;56(1):38-50. doi: 10.1111/jipb.12111. Epub 2013 Nov 20.

31. 牡丹：Transcriptome Comparison Reveals Key Candidate Genes Responsible for the Unusual Reblooming Trait in Tree Peonies. Genes Responsible for Reblooming in Tree Peonies. November 2013 | Volume 8 | Issue 11 | e79996

32. 东南景天：Role of sulfur assimilation pathway in cadmium hyperaccumulation by Sedum alfredii Hance. Ecotoxicology and Environmental Safety. Volume 100, February 2014, Pages 159–165.

33. 山苍子：Identification of appropriate reference genes for normalizing transcript expression by quantitative real‑time PCR in Litsea cubeba. TECHNICAL NOTE. Mol Genet Genomics (2013) 288:727–737, DOI 10.1007/s00438-013-0785-1

34. 木本植物：Heterologous gene silencing induced by tobacco rattle virus (TRV) is efficient for pursuing functional genomics studies in woody plants. ORIGINAL PAPER. Plant Cell Tiss Organ Cult, DOI 10.1007/s11240-013-0393-0

35. 棉花：Analysis of sea-island cotton and upland cotton in response to Verticillium dahliae infection by RNA sequencing. Sun et al. BMC Genomics 2013, 14:852 /1471-2164/14/852.

36. 桃子：Biochemical changes and defence responses during the development of peach gummosis caused by Lasiodiplodia theobromae. Eur J Plant Pathol (2014) 138:195–207, DOI 10.1007/s10658-013-0322-4.

37. 桃子1：Carbohydrate metabolism changes in Prunus persica gummosis infected with Lasiodiplodia theobromae. Phytopathology "First Look" paper • http://dx.doi.org/10.1094/PHYTO-01-13-0025-R • posted 11/27/2013.

38. 海棠：The Malus crabapple transcription factor McMYB10 regulatesanthocyanin biosynthesis during petal coloration. Scientia Horticulturae 166 (2014) 42–49.

39. 海藻：A rapid and sensitive method for field detection of Prorocentrum donghaiense using reverse transcription-coupled loop-mediated isothermal amplification. Harmful Algae 29 (2013) 31–39.

40. 油茶：Establish a cDNA-AFLP Technology System in Camellia oleifera. Molecular Plant Breeding, 2013, Vol.11, No.5, 611-616.

41. 亚洲百合：Transcriptomic analysis of Asiatic lily in the process of vernalization via RNA-seq. Mol Biol Rep. DOI 10.1007/s11033-014-3250-2.

42. 毛泡桐：Dynamic expression of novel and conserved microRNAs and their targets in diploid and tetraploid of Paulownia tomentosa. Biochimie xxx (2014) 1e10.

43. 人参：Cloning and Sequence Analysis Squalene Epoxidase Gene in Panax gin-seng. Journal of Jilin Agricultural University 2014, 36(2): 149-152,17

44. 雪莲：Cloning and Sequence Analysis of rbcs Gene from Sasussured involucrdta Kar. et Kir. Chinese Agricultural Science Bulletin 2014, 30(15): 261-267

45. 柑橘3：Secreted Expression of Cecropin B Gene Enhances Resistance to Xanthomonas axonopodis pv. citri in Transgenic Citrus sinensis‘Tarocco’ Acta Horticulturae Sinica 2014, 41(3): 417–428 http: // www. ahs. ac. cn

46. 菊花：Stem apex detoxification culture markedly improved severalphysiological characters of chrysanthemum ‘YUTAI’. Plant Cell Tiss Organ Cult 2014, DOI 10.1007/s11240-014-0541-1

47. 荞麦和拟南芥：Ectopic expression of FaesAP3, a Fagopyrum esculentum (Polygonaceae) AP3 orthologous gene rescues stamen development in an Arabidopsis ap3 mutant. Gene 2014, 550(2): 200–206

48. 油松：Differential expression of SLOW WALKER2 homologue in ovules of female sterile mutant and fertile clone of Pinus tabulaeformis. Russian Journal of Developmental Biology 2014, 45(2): 78-84

49. 玫瑰花：Precise spatio-temporal modulation of ACC synthase by MPK6 cascade mediates the response of rose flowers to rehydration. The Plant Journal 2014, 79(6): 941–950

50. 棉花和拟南芥：Functional characterization of GhAKT1, a novel Shaker-like K+ channel gene involved in K+ uptake from cotton (Gossypium hirsutum). Gene 2014, 545(1): 61–71

51. 棉花和拟南芥1：Upland Cotton Gene GhFPF1 Confers Promotion of Flowering Time and Shade-Avoidance Responses in Arabidopsis thaliana. PLoS ONE 2014, 9(3): e91869. doi:10.1371/journal.pone.0091869

52. 白杨：Poplar GATA transcription factor PdGNC is capable of regulating chloroplast ultrastructure, photosynthesis, and vegetative growth in Arabidopsis under varying nitrogen levels. Plant Cell Tiss Organ Cult 2014, DOI 10.1007/s11240-014-0536-y

53. 毛果杨：Molecular characterization of the SPL gene family in Populus trichocarpa. BMC Plant Biology 2014, 14: 131

54. 葛根：Molecular cloning and characterization of an isoflavone 7-O-glucosyltransferase from Pueraria lobata. Plant Cell Reports 2014, 33(7), 1173–1185

55. 百合：Cloning and Expression Analysis of Actin Gene（lilyActin）from Lily. Acta Horticulturae Sinica 2013, 40(7): 1318–1326

56. 百合1： Vernalization of Oriental hybrid lily ‘Sorbonne’: changes in physiology metabolic activity and molecular mechanism.Molecular Biology Reports 2014, DOI 10.1007/s11033-014-3545-3

57. 黄鹌菜：Transcriptome Sequencing and De Novo Analysis of Youngia japonica Using the Illumina Platform. PLoS ONE 2014, 9(3): e90636. doi:10.1371/journal.pone.0090636

58. 棉花1：Gibberellin Overproduction Promotes Sucrose Synthase Expression and Secondary Cell Wall Deposition in Cotton Fibers. PLoS ONE 2014, 9(5): e96537. doi:10.1371/journal.pone.0096537

59. 苹果：Low Medium pH Value Enhances Anthocyanin Accumulation in Malus Crabapple Leaves. PLoS ONE 2014, 9(6): e97904. doi:10.1371/journal.pone.0097904

其它公司多糖多酚植物RNA提取试剂盒失败原因和解决方案

很多植物RNA的样品由于含有大量的多糖、多酚、代谢产物、色素等成分，造成RNA提取过程中氧化、褐化、降解、由于植物品种的多样性造成情况更加复杂。手工的CTAB类的方法提取因为时间太长，太繁琐，手工方法不在讨论之列。一直以来没有一款好的试剂盒包括qiagen、promega等进口试剂盒也无法满足科研工作者对植物RNA提取的要求。

下面我们来分析一下植物RNA为什么不能提取成功的原因：

市面上最常见的RNA提取试剂盒无非是两种：第一种：TRIzol改良类方法（包括溶液型的和离心柱型的）、第二种：直接裂解过柱子的方法（离心柱）

第一种试剂盒失败的原因1：RNA市面上面最流行的方法就是Trizol，或者Triol改良，或者Trizol加离心柱一类的改良方法。trizol也就是异硫氰酸胍/苯酚/氯仿原理一步法的方法最适合的对象是动物源性的组织细胞，针对普通多糖多酚低的植物性的材料，TRIzol类原理产品也可以提取。但是多糖、多酚、次级代谢产物丰富的情况下，trizol类方法无法防止多糖多酚对于RNA/DNA分相的干扰，要么残留大量DNA，要么残留大量多糖、多酚或者次级代谢产物，氧化破坏RNA，或者残留这些多糖多酚，色素代谢产物等抑制下游的反转录等反应。限于技术水平的限制，市面上绝大多数的国产厂家是使用trizol的方法进行改良，无论是不是加了离心柱。但是实践证明，改良不能从根本上解决问题。判断是否试剂盒使用这种改良的方法非常简单：是否裂解液含有苯酚的味道和使用氯仿，如果使用到了氯仿就是TRIzol方法的改良。

第二种试剂盒失败的原因：直接裂解过柱子的方法是目前最先进的方法，但是也是技术含量最高的方法。这个方法采用裂解液（不含苯酚，氯仿）直接裂解，RNA/DNA同时过柱子，然后在柱子上面直接分离RNA/DNA，所以，这种方法的优点第一在于，避免了使用trizol在多糖多酚下不能成功分离RNA/DNA的弊端、第二在于，不使用有毒的苯酚氯仿。但是正是因为其技术先进，所以难度很高，国内厂家包括进口公司有两个技术难点一直没有突破。第一，裂解液的成分必须针对去除多糖多酚进行研发添加去多糖多酚，代谢产物成分。否则会同样碰到多糖多酚干扰提取的问题。第二、和trizol原理不同，直接过柱法DNA/RNA同时加到吸附柱上去。如何去除DNA是第一个难点。否则会残留大量DNA。两个技术难点的没有掌握导致了国内公司包括的第二种试剂盒失败。国外公司因为没有掌握第一难点，裂解液里面没有去除多糖多酚成分，所以包括qiagen的盒子也常常不能成功提取植物RNA样品。

本人领衔和研发团队配合经过3年的不断研发改良，针对多糖多酚植物的特点，和这两种试剂盒失败的原因，开发出了EASYspin植物RNA快速提取试剂盒，第一采用直接过柱子方法，彻底抛弃了TRIZOL