Identification of Low Temperature Stress-responsive Genes Regulating Photosynthetic Characteristics in the Leaves of Brassica napus by RNA-Seq

doi:10.13560/j.cnki.biotech.bull.1985.2021-1048

Abstract

Abstract:

In order to clarify the response of photosynthetic characteristics to low temperature stress in Brassica napus L.,using cold-resistant material 17NS and cold-sensitive material NF24 as study object,Illumina HiSeq^TM 2000 platform was applied to have RNA-Seq analysis,the molecular mechanism of B. napus responding to low temperature,such as leaf morphology,photosynthetic parameters,leaf ultrastructure and gene regulation expression pattern treated at -4℃ for 24 h was investigated. The results showed that the photosynthetic pathway and the photosynthesis-antenna protein pathway were significantly enriched in B. napus in response to low temperature stress. In these two pathways,64 genes were specifically expressed,including 8 up-regulated genes and 56 down-regulated genes,most of which were noted in molecular composition and biological process. It was confirmed that photosynthesis was related to leaf wilt,chloroplast number and morphology change,thylakoid structure damage and relative electrolyte permeability increase. qRT-PCR analysis showed that the expression patterns of the first 17 DEGs were consistent with RNA-Seq analysis results,confirming the reliability of RNA-Seq results. BnFd1（LOC106350097）and BnCP26（LOC106440450）were up-regulated in response to cold stress. The amino acid product encoded by BnFd1was the chloroplast ferridoretin 1（BnFd1）,an unstable hydrophilic protein,which were located in the chloroplast and cytoplasm,there was a signal peptide,belonging to the PLNO3136 superfamily and had a conserved Fd1 domain. The amino acid product encoded by BnCP26 was the chlorophyll a-b binding protein CP26（BnCP26）on thylakoid,it was a stable hydrophilic protein located in chloroplast without signal peptide,and belonged to the chlorophyll a_b binding superfamily and had a conserved chlorophyll a-b binding protein domain. Two cold response genes BnFd1 and BnCP26 are successfully identified in B. napus L.,which are closely related to the regulation of photosynthetic characteristics and had conserved interspecies functions.

Key words: Brassica napus L., low temperature stress, differential expressed genes, BnFd1, BnCP26

JIN Jiao-jiao, LIU Zi-gang, MI Wen-bo, XU Ming-xia, ZOU Ya, XU Chun-mei, ZHAO Cai-xia. Identification of Low Temperature Stress-responsive Genes Regulating Photosynthetic Characteristics in the Leaves of Brassica napus by RNA-Seq[J]. Biotechnology Bulletin, 2022, 38(4): 126-142.

Figures/Tables 20

References 44

[1]	姜丽霞, 王萍, 王冬冬, 等. 2013年黑龙江省春季低温特征及对主要农作物播种期的影响[J]. 中国农业气象, 2019, 40(2):114-125.
	Jiang LX, Wang P, Wang DD, et al. Characteristics of low temperature in spring and its effect on crops seeding dates in 2013 in Heilongjiang Province[J]. Chin J Agrometeorology, 2019, 40(2):114-125.
[2]	Lu Q, Li J, Wang J, et al. Exploration of a mechanism for the production of highly unsaturated fatty acids in Scenedesmus sp. at low temperature grown on oil crop residue based medium[J]. Bioresour Technol, 2017, 244(pt 1):542-551. doi: 10.1016/j.biortech.2017.08.005 URL
[3]	刘文彬. 低温冷害对农作物生长发育的影响[J]. 黑龙江科技信息, 2011(19):204.
	Liu WB. Effects of chilling injury on crop growth and developmen[J]. Heilongjiang Sci Technol Inf, 2011(19):204.
[4]	Dahanayake SR, Galwey NW. Effects of interactions between low-temperature treatments, gibberellin(GA3)and photoperiod on flowering and stem height of spring rape(Brassica napus var. annua)[J]. Ann Bot, 1999, 84(3):321-327. doi: 10.1006/anbo.1999.0920 URL
[5]	涂玉琴, 戴兴临. 花期低温阴雨对甘蓝型油菜产量和种子含油量的影响[J]. 中国油料作物学报, 2011, 33(5):470-475.
	Tu YQ, Dai XL. Effects of continuous low temperature overcast and rainy weather on yield and oil content of Brassica napus during flowering stage[J]. Chin J Oil Crop Sci, 2011, 33(5):470-475.
[6]	卢坤, 张琳, 曲存民, 等. 利用RNA-Seq鉴定甘蓝型油菜叶片干旱胁迫应答基因[J]. 中国农业科学, 2015, 48(4):630-645.
	Lu K, Zhang L, Qu CM, et al. Identification of drought stress-responsive genes in leaves of Brassica napus by RNA sequencing[J]. Sci Agric Sin, 2015, 48(4):630-645.
[7]	陈永川, 张林, 杨红艳. 低温冷害对农作物的危害及其防御措施[J]. 现代农业科技, 2011(4):307.
	Chen YC, Zhang L, Yang HY. The harm of low temperature chilling injury to crops and its preventive measures[J]. Mod Agric Sci Technol, 2011(4):307.
[8]	何跃君, 薛立, 任向荣, 等. 低温胁迫对六种苗木生理特性的影响[J]. 生态学杂志, 2008, 27(4):524-531.
	He YJ, Xue L, Ren XR, et al. Effects of low temperature stress on physiological characteristics of six tree species seedlings[J]. Chin J Ecol, 2008, 27(4):524-531.
[9]	许楠, 孙广玉. 低温锻炼后桑树幼苗光合作用和抗氧化酶对冷胁迫的响应[J]. 应用生态学报, 2009, 20(4):761-766.
	Xu N, Sun GY. Responses of mulberry seedlings photosynjournal and antioxidant enzymes to chilling stress after low-temperature acclimation[J]. Chin J Appl Ecol, 2009, 20(4):761-766.
[10]	Oquist G. Effects of low temperature on photosynjournal[J]. Plant Cell Environ, 1983, 6(4):281-300.
[11]	Zhuang WF, Wu XJ, Yang M, et al. Influence of low-temperature stress on photosynthetic traits in maize seedlings[J]. J Northeast Agric Univ:Engl Ed, 2013, 20(3):1-5.
[12]	Holá D, Langrová K, Kočová M, et al. Photosynthetic parameters of maize(Zea mays L.)inbred lines and F1 hybrids:their different response to, and recovery from rapid or gradual onset of low-temperature stress[J]. Photosynthetica, 2003, 41(3):429-442. doi: 10.1023/B:PHOT.0000015468.67892.9c URL
[13]	陈建林, 查丁石, 吴雪霞. 低温胁迫对茄子幼苗生理生化的影响[J]. 中国蔬菜, 2006(11):21-23.
	Chen JL, Zha DS, Wu XX. Effects of low temperature stress on physiology and biochemistry of eggplant seedlings[J]. China Veg, 2006(11):21-23.
[14]	李晓靖, 崔海军. 低温胁迫下植物光合生理研究进展[J]. 山东林业科技, 2018, 48(6):90-94.
	Li XJ, Cui HJ. Research progress on the physiological response of plants to environmental stress[J]. J Shandong For Sci Technol, 2018, 48(6):90-94.
[15]	Astakhova NV, Popov VN, Selivanov AA, et al. Reorganization of chloroplast ultrastructure associated with low-temperature hardening of Arabidopsis plants[J]. Russ J Plant Physiol, 2014, 61(6):744-750. doi: 10.1134/S102144371406003X URL
[16]	Arthikala MK, Sánchez-López R, Nava N, et al. RbohB, a Phaseolus vulgaris NADPH oxidase gene, enhances symbiosome number, bacteroid size, and nitrogen fixation in nodules and impairs mycorrhizal colonization[J]. New Phytol, 2014, 202(3):886-900. doi: 10.1111/nph.12714 URL
[17]	Shi Y, Gong Z. One SNP in COLD1 determines cold tolerance during rice domestication[J]. J Genet Genomics, 2015, 42(4):133-134. doi: 10.1016/j.jgg.2015.03.007 URL
[18]	杨刚, 史鹏辉, 孙万仓, 等. 白菜型冬油菜质外体抗冻蛋白研究[J]. 中国生态农业学报, 2016, 24(2):210-217.
	Yang G, Shi PH, Sun WC, et al. Study on apoplast anti-freeze proteins in winter turnip rape(Brassica rape L.)[J]. Chin J Eco Agric, 2016, 24(2):210-217.
[19]	Huang Q, Qian X, Jiang T, et al. Effect of eugenol fumigation treatment on chilling injury and CBF gene expression in eggplant fruit during cold storage[J]. Food Chem, 2019, 292:143-150. doi: 10.1016/j.foodchem.2019.04.048 URL
[20]	王政, 姚银安, 张志燕, 等. 甘蓝型油菜几丁质酶基因BnCHB4环境信号响应分析[J]. 生物学杂志, 2012, 29(6):32-34, 38.
	Wang Z, Yao YA, Zhang ZY, et al. Response to environmental cues of BnCHB4 from Brassica napus[J]. J Biol, 2012, 29(6):32-34, 38.
[21]	张腾国, 罗丹瑜, 郭艳峰, 等. 油菜RbohB基因的克隆及逆境响应表达分析[J]. 植物研究, 2017, 37(5):751-760.
	Zhang TG, Luo DY, Guo YF, et al. Cloning and expression analysis of RbohB gene in Brassica campestris L[J]. Bull Bot Res, 2017, 37(5):751-760.
[22]	杨佳, 曹黎明, 周继华, 等. 水稻耐低温基因COLD1功能标记的开发及应用[J]. 分子植物育种, 2019, 17(18):6028-6032.
	Yang J, Cao LM, Zhou JH, et al. Development and application of functional marker of chilling tolerance gene, COLD1, in rice[J]. Mol Plant Breed, 2019, 17(18):6028-6032.
[23]	王冰, 王勤方, 唐天向, 等. 转录组分析挖掘油菜耐旱基因[J]. 基因组学与应用生物学, 2018, 37(11):4775-4786.
	Wang B, Wang QF, Tang TX, et al. Discovery of drought-tolerant genes in Brassica napus by transcriptome analysis[J]. Genom Appl Biol, 2018, 37(11):4775-4786.
[24]	Horvath DP, McLarney BK, Thomashow MF. Regulation of Arabidopsis thaliana L. (heyn)cor78 in response to low temperature[J]. Plant Physiol, 1993, 103(4):1047-1053. pmid: 8290624
[25]	Zhang Z, Li J, Li F, et al. OsMAPK3 phosphorylates OsbHLH002/OsICE1 and inhibits its ubiquitination to activate OsTPP1 and enhances rice chilling tolerance[J]. Dev Cell, 2017, 43(6):731-743.e5. doi: 10.1016/j.devcel.2017.11.016 URL
[26]	陈珊, 王晓晨, 邝健飞, 等. 黄瓜果实耐冷性与CsHSFs基因表达关系的研究[J]. 华南农业大学学报, 2015, 36(5):85-91.
	Chen S, Wang XC, Kuang JF, et al. The expressions of CsHSFs gene in relation to chilling tolerance of cucumber fruits[J]. J South China Agric Univ, 2015, 36(5):85-91.
[27]	Talanova VV, Titov AF, Topchieva LV, et al. Effect of stress factors on expression of the gene encoding a CBF transcription factor in cucumber plants[J]. Dokl Biol Sci, 2008, 423:419-421. doi: 10.1134/S001249660806015X URL
[28]	刘自刚, 张长生, 孙万仓, 等. 不同生态区冬前低温下白菜型冬油菜不同抗寒品种(系)的比较[J]. 作物学报, 2014, 40(2):346-354.
	Liu ZG, Zhang CS, Sun WC, et al. Comparison of winter rapeseed varieties(lines)with different cold resistance planted in the northern-extending regions in China under low temperature before winter[J]. Acta Agron Sin, 2014, 40(2):346-354. doi: 10.3724/SP.J.1006.2014.00346 URL
[29]	邹琦. 植物生理学实验指导[M]. 北京: 中国农业出版社, 2000.
	Zou Q. Experimental Guidance on Plant Physiology[M]. Beijing: China Agricultural Press, 2000.
[30]	张屹清, 林路友, 路争. 严格厌氧菌铁氧还蛋白的研究进展[J]. 微生物学报, 2021. DOI: 10.13343/j.cnki.wsxb,20210146. doi: 10.13343/j.cnki.wsxb,20210146
	Zhang YQ, Lin LY, Lu Z. Advances of ferredoxins from strictly anaerobic bacteria[J]. Acta Microbiol Sin, 2021. DOI: 10.13343/j.cnki.wsxb,20210146. doi: 10.13343/j.cnki.wsxb,20210146
[31]	Farquhar GD, Sharkey TD. Stomatal conductance and photosynjournal[J]. Annu Rev Plant Physiol, 1982, 33(1):317-345. doi: 10.1146/annurev.pp.33.060182.001533 URL
[32]	杨碧云, 叶丽萍, 钟凤林, 等. 低温处理对紫色小白菜品质及光合特性的影响[J]. 安徽农业大学学报, 2019, 46(1):173-180.
	Yang BY, Ye LP, Zhong FL, et al. Effects of low-temperature stress on the quality and photosynthetic characteristics of purple cabbage[J]. J Anhui Agric Univ, 2019, 46(1):173-180.
[33]	武辉, 戴海芳, 张巨松, 等. 棉花幼苗叶片光合特性对低温胁迫及恢复处理的响应[J]. 植物生态学报, 2014, 38(10):1124-1134. doi: 10.3724/SP.J.1258.2014.00107
	Wu H, Dai HF, Zhang JS, et al. Responses of photosynthetic characteristics to low temperature stress and recovery treatment in cotton seedling leaves[J]. Chin J Plant Ecol, 2014, 38(10):1124-1134. doi: 10.3724/SP.J.1258.2014.00107 URL
[34]	许耀照, 张芬琴, 陈修斌, 等. 低温胁迫对彩椒幼苗生长指标及光合特性的影响[J]. 山西农业大学学报:自然科学版, 2019, 39(1):68-72.
	Xu YZ, Zhang FQ, Chen XB, et al. Effects of low temperature stress on growth index and photosynthetic characteristics of pepper seedlings[J]. J Shanxi Agric Univ:Nat Sci Ed, 2019, 39(1):68-72.
[35]	田景花, 王红霞, 张志华, 等. 低温逆境对不同核桃品种抗氧化系统及超微结构的影响[J]. 应用生态学报, 2015, 26(5):1320-1326.
	Tian JH, Wang HX, Zhang ZH, et al. Effects of chilling stress on antioxidant system and ultrastructure of walnut cultivars[J]. Chin J Appl Ecol, 2015, 26(5):1320-1326.
[36]	陈颖, 陈昕, 汪南阳, 等. 低温胁迫下西番莲叶片的生理反应及超微结构变化[J]. 西北植物学报, 2012, 32(3):532-539.
	Chen Y, Chen X, Wang NY, et al. Physiological response to cold stress and cell ultra-structure changes in Passiflora edulis leaves[J]. Acta Bot Boreali Occidentalia Sin, 2012, 32(3):532-539.
[37]	郑国华, 张贺英, 钟秀容. 低温胁迫下枇杷叶片细胞超微结构及膜透性和保护酶活性的变化[J]. 中国生态农业学报, 2009, 17(4):739-745.
	Zheng GH, Zhang HY, Zhong XR. Changes in cell ultra-structure, membrane permeability and protective enzyme activity in Eriobotrya japonica Lindl. leaves under cold stress[J]. Chin J Eco Agric, 2009, 17(4):739-745. doi: 10.3724/SP.J.1011.2009.00739 URL
[38]	梁李宏, 梅新, 林锋, 等. 低温胁迫对腰果幼苗叶片组织结构和生理指标的影响[J]. 生态环境学报, 2009, 18(1):317-320.
	Liang LH, Mei X, Lin F, et al. Effect of low temperature stress on tissue structure and physiological index of cashew young leaves[J]. Ecol Environ Sci, 2009, 18(1):317-320.
[39]	王晨光, 王希, 苍晶, 等. 低温胁迫对水稻幼苗抗冷性的影响[J]. 东北农业大学学报, 2004, 35(2):205-207.
	Wang CG, Wang X, Cang J, et al. Effect of low-temperature stress on cold-resistance ability of rice seedlings[J]. J Northeast Agric Univ, 2004, 35(2):205-207.
[40]	付琳, 厉辉, 张文山, 等. 低温胁迫对不同拔节进程冬小麦光合特性的影响[J]. 新农业, 2021(13):69-71.
	Fu L, Li H, Zhang WS, et al. Effects of low temperature stress on photosynthetic characteristics of winter wheat at different jointing stages[J]. New Agri, 2021(13):69-71.
[41]	董杰, 张昊, 张芃芃. 集胞藻PCC 6803类铁氧还蛋白Slr1205的功能研究[J]. 生物技术进展, 2021, 11(1):69-78.
	Dong J, Zhang H, Zhang PP. Functional study on ferredoxin-like protein Slr1205 in Synechocystis sp. PCC 6803[J]. Curr Biotechnol, 2021, 11(1):69-78.
[42]	吴晓佩. 文心兰离体培养优化及转化铁氧还蛋白基因研究[D]. 福州:福建农林大学, 2017.
	Wu XP. Optimization of Oncidium in vitro culture system and transformation ananlysis of ferredoxin genes[D]. Fuzhou:Fujian Agriculture and Forestry University, 2017.
[43]	Marin A, Passarini F, Croce R, et al. Energy transfer pathways in the CP24 and CP26 antenna complexes of higher plant photosystem II:a comparative study[J]. Biophys J, 2010, 99(12):4056-4065. doi: 10.1016/j.bpj.2010.10.034 URL
[44]	Cazzaniga S, Kim M, Bellamoli F, et al. Photosystem II antenna complexes CP26 and CP29 are essential for nonphotochemical quenching in Chlamydomonas reinhardtii[J]. Plant Cell Environ, 2020, 43(2):496-509. doi: 10.1111/pce.13680

KEGG通路 KEGG pathway	基因号 Gene ID	引物序列 Sequence of primer（5'-3'）	扩增长度 Amplification length/bp	扩增效率 Amplification efficiency/%
P	ncbi_106366341	AGATCCTAATGCCACCATCATC（F）	100	101.80
	ncbi_106366341	GTAGTCCTCATGCTCCTCAAAG（R）	100	101.80
	ncbi_106360470	CTCCTTACAACCCACTTCAGAG（F）	100	96.80
	ncbi_106360470	GAGCCACCTCCAAGAATCAA（R）	100	96.80
	ncbi_106400646	CTACCGACAAGAAGTCCATCAC（F）	102	88.28
	ncbi_106400646	AGGCAGTCTCACCGAAGTA（R）	102	88.28
	ncbi_106430073	GCAGAGGCAAGGATCAAAGA（F）	99	93.16
	ncbi_106430073	GTAAGGACGACGGAGGAAATAAG（R）	99	93.16
	ncbi_106381576	GCAGAGGCAAGGATCAAAGA（F）	99	100.09
	ncbi_106381576	GTAAGGACGACGGAGGAAATAAG（R）	99	100.09
	ncbi_106401683	CCGAGATTTGCTCTCCTTATCC（F）	119	103.61
	ncbi_106401683	CCGACGTATCTTTCACGTACTC（R）	119	103.61
	ncbi_106405529	CTCCTTACAACCCACTTCAGAG（F）	100	95.07
	ncbi_106405529	GAGCCTCCTCCAAGAATCAAA（R）	100	95.07
	ncbi_106350097	CATAACACCAGATGGAGAGAAGG（F）	149	102.17
	ncbi_106350097	CAACCGAGCCAGAGACTATTT（R）	149	102.17
PAP	ncbi_106371874	TAGCTGGAGATTACGGGTTTG（F）	143	122.90
	ncbi_106371874	GGTACGAAGTAGGTCGGTTAAG（R）	143	122.90
	ncbi_106382137	CTCGGAAACCCTAACTTGATCC（F）	100	101.97
	ncbi_106382137	CACCTCCAATTCTGTACCCTTC（R）	100	101.97
	ncbi_106439795	AACCGTGAGCTCGAAGTAATC（F）	100	98.06
	ncbi_106439795	CTCCGAATTTGACTCCGTTCT（R	100	98.06
	ncbi_106349237	AACCGTGAGCTCGAAGTAATC（F）	131	102.84
	ncbi_106349237	ATCTGAGAACCTGCCTTGAAC（R）	131	102.84
	ncbi_106396209	GAAGGTGGGCTCGACTATTT（F）	112	94.26
	ncbi_106396209	CGACTCTGTAACCCTCAACAG（R）	112	94.26
	ncbi_106447496	GAAGGAGGGCTTGACTACTTG（F）	99	103.51
	ncbi_106447496	CTCAACAGCTCCCATGAGAAT（R）	99	103.51
	ncbi_106447497	AAACGGAAGGTTGGCTATGT（F）	96	100.88
	ncbi_106447497	GCCAAATGGTCAGCAAGATTC（R）	96	100.88
	ncbi_106421061	CTGTCAAGCAAGGAGCAAAC（F）	102	97.35
	ncbi_106421061	CAAGAGGGTCGAATCCATAGTC（R）	102	97.35
	ncbi_106440450	AACCTTGTTCTGGCTGTAGTT（F）	98	93.57
	ncbi_106440450	GGGTGTAGCTTGTCCTCAAA（R）	98	93.57
RG	Bnactin-F	TCCATCCATCGTCCACAG		106.80
RG	Bnactin-R	GCATCATCACAAGCATCCTT		106.80

KEGG通路 KEGG pathway	基因号 Gene ID	引物序列 Sequence of primer（5'-3'）	扩增长度 Amplification length/bp	扩增效率 Amplification efficiency/%
P	ncbi_106366341	AGATCCTAATGCCACCATCATC（F）	100	101.80
	ncbi_106366341	GTAGTCCTCATGCTCCTCAAAG（R）	100	101.80
	ncbi_106360470	CTCCTTACAACCCACTTCAGAG（F）	100	96.80
	ncbi_106360470	GAGCCACCTCCAAGAATCAA（R）	100	96.80
	ncbi_106400646	CTACCGACAAGAAGTCCATCAC（F）	102	88.28
	ncbi_106400646	AGGCAGTCTCACCGAAGTA（R）	102	88.28
	ncbi_106430073	GCAGAGGCAAGGATCAAAGA（F）	99	93.16
	ncbi_106430073	GTAAGGACGACGGAGGAAATAAG（R）	99	93.16
	ncbi_106381576	GCAGAGGCAAGGATCAAAGA（F）	99	100.09
	ncbi_106381576	GTAAGGACGACGGAGGAAATAAG（R）	99	100.09
	ncbi_106401683	CCGAGATTTGCTCTCCTTATCC（F）	119	103.61
	ncbi_106401683	CCGACGTATCTTTCACGTACTC（R）	119	103.61
	ncbi_106405529	CTCCTTACAACCCACTTCAGAG（F）	100	95.07
	ncbi_106405529	GAGCCTCCTCCAAGAATCAAA（R）	100	95.07
	ncbi_106350097	CATAACACCAGATGGAGAGAAGG（F）	149	102.17
	ncbi_106350097	CAACCGAGCCAGAGACTATTT（R）	149	102.17
PAP	ncbi_106371874	TAGCTGGAGATTACGGGTTTG（F）	143	122.90
	ncbi_106371874	GGTACGAAGTAGGTCGGTTAAG（R）	143	122.90
	ncbi_106382137	CTCGGAAACCCTAACTTGATCC（F）	100	101.97
	ncbi_106382137	CACCTCCAATTCTGTACCCTTC（R）	100	101.97
	ncbi_106439795	AACCGTGAGCTCGAAGTAATC（F）	100	98.06
	ncbi_106439795	CTCCGAATTTGACTCCGTTCT（R	100	98.06
	ncbi_106349237	AACCGTGAGCTCGAAGTAATC（F）	131	102.84
	ncbi_106349237	ATCTGAGAACCTGCCTTGAAC（R）	131	102.84
	ncbi_106396209	GAAGGTGGGCTCGACTATTT（F）	112	94.26
	ncbi_106396209	CGACTCTGTAACCCTCAACAG（R）	112	94.26
	ncbi_106447496	GAAGGAGGGCTTGACTACTTG（F）	99	103.51
	ncbi_106447496	CTCAACAGCTCCCATGAGAAT（R）	99	103.51
	ncbi_106447497	AAACGGAAGGTTGGCTATGT（F）	96	100.88
	ncbi_106447497	GCCAAATGGTCAGCAAGATTC（R）	96	100.88
	ncbi_106421061	CTGTCAAGCAAGGAGCAAAC（F）	102	97.35
	ncbi_106421061	CAAGAGGGTCGAATCCATAGTC（R）	102	97.35
	ncbi_106440450	AACCTTGTTCTGGCTGTAGTT（F）	98	93.57
	ncbi_106440450	GGGTGTAGCTTGTCCTCAAA（R）	98	93.57
RG	Bnactin-F	TCCATCCATCGTCCACAG		106.80
RG	Bnactin-R	GCATCATCACAAGCATCCTT		106.80

反应物 Reagent	体积Volume/μL
SYBR^R Premix Ex Taq ^TM II	10
Forward primer	0.8
Reverse primer	0.8
ROX Reference Dye II	0.4
Template cDNA	2
ddH₂O	6

反应物 Reagent	体积Volume/μL
SYBR^R Premix Ex Taq ^TM II	10
Forward primer	0.8
Reverse primer	0.8
ROX Reference Dye II	0.4
Template cDNA	2
ddH₂O	6

组合 Paired comparison	显著上调差异基因数 Significantly up-regulated differential genes	显著下调差异基因数 Significantly down-regulated differential genes	总差异基因数 Significantly different number of genes
NF24 t0 - NF24 t	12 583	24 414	36 997
17NS t0 - 17NS t	11 710	23 141	34 851
NF24 t0-17NS t0	10 708	8 270	18 978
NF24 t - 17NS t	11 447	10 572	22 019