杨雨璋;周贝贝;李民吉;李兴亮;张军科;张强;魏钦平;

• 1:北京市林业果树科学研究院·农业农村部华北地区园艺作物生物学与种质创制重点实验室·北京市落叶果树工程技术研究中心

摘要(Abstract)：

【目的】探讨培养基大量元素组分和配比对苹果矮化砧木‘SH6’组培继代的影响,为提高组培快繁效率、建立快繁技术提供理论依据。【方法】采用五因素二次回归正交旋转设计(1/2实施)方法,以MS培养基大量元素KNO_3、NH_4NO_3、MgSO_4·7H_2O、KH_2PO_4与CaCl_2·2H_2O为自变量,接种后35 d的组培苗继代系数和幼苗高度为因变量。【结果】单因子效应分析表明,大量元素影响苹果矮化砧木‘SH6’继代系数的主次顺序是KNO_3> NH_4NO_3> MgSO_4·7H_2O>KH_2PO_4> CaCl_2·2H_2O,影响幼苗高度的主次顺序是KNO_3> NH_4NO_3> CaCl_2·2H_2O> MgSO_4·7H_2O> KH_2PO_4。因子交互效应表明,继代系数受NH_4NO_3与CaCl_2·2H_2O交互效应的影响;幼苗高度受KNO_3与NH_4NO_3、KNO_3与KH_2PO_4、KNO_3与CaCl_2·2H_2O交互效应的影响。配方优化的研究表明,理论预测与实验验证的结果无显著差异。【结论】MS培养基的大量元素影响苹果矮化砧木‘SH6’的继代系数和幼苗高度,KNO_3与NH_4NO_3是主要影响因素。大量元素优化配方为:KNO_31 535 mg·L~(-1)、NH_4NO_31 430 mg·L~(-1)、MgSO_4·7H_2O 310 mg·L~(-1)、KH_2PO_4172 mg·L~(-1)与CaCl_2·2H_2O 445 mg·L~(-1),培养35 d继代系数4.2,幼苗高度2.6 cm。

关键词(KeyWords)： 苹果矮化砧木;‘SH6’;组织培养;大量元素;二次回归正交旋转设计;配方优化

Abstract:

Keywords:

基金项目(Foundation):

作者(Author): 杨雨璋;周贝贝;李民吉;李兴亮;张军科;张强;魏钦平;

Email:

DOI: 10.13925/j.cnki.gsxb.20190213

参考文献(References)：

• [1]白海霞.苹果M26自根砧树的生产效应分析[D].杨凌：西北农林科技大学，2008.BAI Haixia. Analysis on production of dwarfing self-root stock apple tree M26[D]. Yangling:Northwest A&F University,2008.
• [2]邓丰产，马锋旺.苹果矮化自根砧嫁接苗繁育技术研究[J].园艺学报，2012,39(7):1353-1358.DENG Fengchan,MA Fengwang. Propagation studies of selfrooted dwarf grafted apple trees[J]. Acta Horticulturae Sinica,2012,39(7):1353-1358.
• [3]李民吉，张强，李兴亮，周贝贝，杨雨璋，周佳，张军科，魏钦平. SH6矮化中间砧‘富士’苹果不同树形对树体生长和果实产量、品质的影响[J].中国农业科学，2017,50(19):3789-3796.LI Minji,ZHANG Qiang,LI Xingliang,ZHOU Beibei,YANG Yuzhang,ZHOU Jia,ZHANG Junke,WEI Qinping. Effect of three different tree shapes on growth,yield and fruit quality of‘fuji’apple trees on dwarfing interstocks[J]. Scientia Agricultura Sinica,2017,50(19):3789-3796.
• [4] BHATTI S,JHA G. Current trends and future prospects of biotechnological interventions through tissue culture in apple[J].Plant Cell Reports,2010,29(11):1215-1225.
• [5] DUBRANSZKI J,da SILVA J A T. Micropropagation of appleA review[J]. Biotechnology Advances,2010,28(4):462-488.
• [6] HOHNLE M K,WEBER G. Efficient adventitious shoot formation of leaf segments of in vitro propagated shoots of the apple rootstock M.9/T337[J]. European Journal of Horticultural Science,2010,75(3):128-131.
• [7] DASTJERD Z H,JABBARZADEH Z,MARANDI R J. Interaction effects of chitosan,benzyladenine,and gibberellic acid on in vitro proliferation of M26 apple rootstock[J]. Horticulture Environment and Biotechnology,2013,54(6):538-547.
• [8] ZHANH X,QIN Y,LIANG D,ZOU Y J,MA F W. Enhancement of in vitro shoot regeneration from leaf explants of apple rootstock G.41[J]. In Vitro Cellular&Developmental BiologyPlant,2014,50(2):263-270.
• [9] YANCHEVA S D,GOLUBOWICZ S,FISHER E,LEV-YADUN S,FLAISHMAN M A. Auxin type and timing of application determine the activation of the developmental program during in vitro organogenesis in apple[J]. Plant Science,2003,165(2):299-309.
• [10] AMIRI E M,ELAHINIA A. Optimization of medium composition for apple rootstocks[J]. African Journal of Biotechnology,2011,10(18):3594-3601.
• [11] TABART J,FRANCK T,KEVERS C,DOMMES J. Effect of polyamines and polyamine precursors on hyperhydricity in micropropagated apple shoots[J]. Plant Cell Tissue and Organ Culture,2015,120(1):11-18.
• [12]邵开基，章德明，陈霜莹，赵艳华，戴桂林，牛自勉. SH系苹果矮化砧茎尖组培快繁技术研究[J].山西农业科学，1995,23(1):14-18.SHAO Kaiji,ZHANG Deming,CHEN Shuangying,ZHAO Yanhua,DAI Guilin,NIU Zimian. A study on rapid propagation technique through stem tip tissue culture of SH line apple dwarf stock[J]. Journal of Shanxi Agricultural Sciences,1995,23(1):14-18.
• [13]周莉.苹果矮化砧木离体培养和快繁体系建立与优化[D].杨凌：西北农林科技大学，2017.ZHOU Li. The foundation and optimization of vitro culture and rapid propagation system of apple dwarfing rootstock[D]. Yang ling:Northwest A&F University,2017.
• [14]肖莉，张国权.回归旋转正交设计的应用及其SAS实施[J].广东交通职业技术学院学报，2009,8(3):81-84.XIAO Li,ZHANG Guoquan. The application of regression rotating orthogonal design and its SAS program[J]. Journal of Guang Dong Communications Polytechnic,2009,8(3):81-84.
• [15] MURASHIGE T,SKOOG F. A revised medium for rapid growth and bio assays with tobacco tissue cultures[J]. Physiologia Plantarum,1962,15(3):473-497.
• [16]杨艳敏，魏永祥，刘成，张舵，王兴东，刘友春.苹果矮化砧木组织培养及脱病毒技术[J].北方园艺，2016(4):107-112.YANG Yanmin,WEI Yongxiang,LIU Cheng,ZHANG Duo,WANG Xingdong,LIU Youchun. Tissue culture and virus elimination technique for apple dwarf rootstock[J]. Northern Horticulture,2016(4):107-112.
• [17]谢璇，许轲，谢闽新，朱元娣.苹果茎尖培养快繁体系的优化[J].植物生理学报，2015,51(12):2152-2156.XIE Xuan,XU Ke,XIE Minxin,ZHU Yuandi. Optimization of rapid micropropagation system of apple meristem-tip culture[J].Plant Physiology Journal,2015,51(12):2152-2156.
• [18] THORPE T A. History of plant tissue culture[J]. Molecular Biotechnology,2007,37(2):169-180.
• [19]上官新晨，蒋艳，米丽雪，陈继光，张清峰，邹利. 5种大量元素对青钱柳愈伤组织生长及黄酮类化合物积累的影响[J].江西农业大学学报，2011,33(3):502-507.SHANGGUAN Xinchen,JIANG Yan,MI Lixue,CHEN Jiguang,ZHANG Qingfeng,ZOU Li. Effects of five macronutrients on the growth and flavonoid accumulation of Cyclocarya paliurus callus[J]. Acta Agriculturae Universitatis Jiangxiensis,2011,33(3):502-507.
• [20] NOWAK B,MICZYNSKI K,HUDY L. The effect of total inorganic nitrogen and the balance between its ionic forms on adventitious bud formation and callus growth of‘Wegierka Zwykla’plum(Prunus domestica L.)[J]. Acta Physiologiae Plantarum,2007,29(5):479-484.
• [21] SUN Q R,SUN M J,SUN H Y,BELL R L,LI L G,ZHANG W,TAO J H. Comparative organogenic response of six clonal apple rootstock cultivars[J]. HortScience,2016,51(3):272-278.
• [22] CHAILLOU S,VESSEY J K,MOROT-GAUDRY C D,RAPER J R,HENRY L T,BOUTIN J P. Expression of characteristics of ammonium nutrition as affected by pH of the root medium[J].Journal of Experimental Botany,1991,42(2):189-196.
• [23] RAMAGE C M,WILLIAMS R R. Mineral nutrition and plant morphogenesis[J]. In Vitro Cellular&Developmental BiologyPlant,2002,38(2):116-124.
• [24] NIEDZ R P,EVENS T J. The effects of nitrogen and potassium nutrition on the growth of nonembryogenic and embryogenic tissue of sweet orange(Citrus sinensis(L.)Osbeck)[J]. BMC Plant Biology,2008,8(1):126.
• [25] WALCH-LIU P,NEUMANN G,BANGERTH F,ENGELS G.Rapid effects of nitrogen form on leaf morphogenesis[J]. Journal of Experimental Botany,2000,51(343):227-237.
• [26] BRITTO D T,KRONZUCKER H J. NH4+toxicity in higher plants:a critical review[J]. Journal of Plant Physiology,2002,159(6):567-584.
• [27] CASTANEDA-MENDEZ O,OGAWA S,MEDINA A,CHAVARRIAGA P,GOMEZ SELVARAJ M. A simple hydroponic hardening system and the effect of nitrogen source on the acclimation of in vitro cassava(Manihot esculenta Crantz)[J]. In Vitro Cellular&Developmental Biology-Plant,2017,53(2):75-85.
• [28] PREDIERI S,FASOLO FABBRI MALAVASI F. High-frequency shoot regeneration from leaves of the apple rootstock M26(Malus pumila Mill.)[J]. Plant Cell,Tissue and Organ Culture,1989,17(2):133-142.
• [29] EL-HAWAZ R F,BRIDGES W C,ADELBERG J W. In vitro growth of Curcuma longa L. in response to five mineral elements and plant density in Fed-Batch culture systems[J]. PLoS One,2015,10(4):e0118912.
• [30] SCHENK N,HSIAO K C,BORNMAN C H. Avoidance of precipitation and carbohydrate breakdown in autoclaved plant tissue culture media[J]. Plant Cell Reports,1991,10(3):115-119.
• [31] GRIBBLE K,CONROY J P,HOLFORD P,MILHAM P J. In vitro uptake of minerals by Gypsophila paniculata and hybrid eucalypts,and relevance to media mineral formulation[J]. Australian Journal of Botany,2002,50(6):713-723.
• [32] JANSEN M A K,BOOIJ H,SCHEL J H N,de VRIES S C. Calcium increases the yield of somatic embryos in carrot embryogenicsuspensioncultures[J].PlantCellReports,1990,9(4):221-223.
• [33] SOTIROPOULOS T E. Effect of NaCl and CaCl2on growth and contents of minerals,chlorophyll,proline and sugars in the apple rootstock M4 cultured in vitro[J]. Biologia Plantarum,2007,51(1):177-180.
• [34] SCHEIBLE W R,LAUERER M,SCHULZE E D,CABOCHE M,STITT M. Accumulation of nitrate in the shoot acts as a signal to regulate shoot-root allocation in tobacco[J]. The Plant Journal,1997,11(4):671-691.
• [35] MARTIN S M,ROSE D. Growth of plant cell(Ipomoea)suspension cultures at controlled pH levels[J]. Canadian Journal of Botany,1976,54(11):1264-1270.
• [36] REED B M,WADA S,DENOMA J,NIEDZ R P. Improving in vitro mineral nutrition for diverse pear germplasm[J]. In Vitro Cellular&Developmental Biology-Plant,2013,49(3):343-355.
• [37] SOTIROPOULOS T,MOUHTARIDOU G,THOMIDIS T,TSIRAKOGLOU V,DIMASSI K,THERIOS I. Effects of different N-sources on growth,nutritional status,chlorophyll content,and photosynthetic parameters of shoots of the apple rootstock MM 106 cultured in vitro[J]. Biologia Plantarum,2005,49(2):297-299.
• [38] GRIMES H D,HODGES T K. The inorganic NO3-:NH4+ratio influences plant regeneration and auxin sensitivity in primary callus derived from immature embryos of indica rice(Oryza sativa L.)[J]. Journal of Plant Physiology,1990,136(3):362-367.
• [39] PRAVEEN N,MURTHY H N. Withanolide a production from Withania somnifera hairy root cultures with improved growth by altering the concentrations of macro elements and nitrogen source in the medium[J]. Acta Physiologiae Plantarum,2013,35(3):811-816.
• [40] MARTIN S M,ROSE D,HUI V. Growth of plant cell suspension cultures with ammonium as the sole source of nitrogen[J].Canadian Journal of Botany,1977,55(22):2838-2843.
• [41] CHOI J M,LATIGUI A,LEE C W. Growth and nutrient uptake responses of‘Seolhyang’strawberry to various ratios of ammonium to nitrate nitrogen in nutrient solution culture using inert media[J]. African Journal of Biotechnology,2011,10(59):12567-12574.
• [42] KOVALCHUK I Y,MUKHITDINOVA Z,TURDIYEV T,MADIYEVA G,AKIN M,EYDURAN E,REED B M. Nitrogen ions and nitrogen ion proportions impact the growth of apricot(Prunus armeniaca)shoot cultures[J]. Plant Cell,Tissue and Organ Culture(PCTOC),2018,133(2):263-273.
• [43] RUGINI E,MUGANU M. A novel strategy for the induction and maintenance of shoot regeneration from callus derived from established shoots of apple[Malus×domestica Borkh.] cv. Golden Delicious[J]. Plant Cell Reports,1998,17(6):581-585.
• [44]谢利娟，韩蕾，钱永强，李晓东，李永红.爆仗竹组培快繁6-BA与NAA组合浓度配比优化分析[J].核农学报，2005,19(3):181-185.XIE Lijuan,HAN Lei,QIAN Yongqiang,LI Xiaodong,LI Yonghong. Optimization of the concentration of 6-BA and NAA in the rapid propagation of Russelia equisetiformis[J]. Journal of Nuclear Agricultural Sciences,2005,19(3):181-185.
• [45]韩秀慧，尹伟伦，王华芳.二次回归正交设计在微型月季组织培养中的应用[J].林业科学，2004,40(4):189-192.HAN Xiuhui,YIN Weilun,WANG Huafang. Application of quadratic regressive factorial experiment to in vitro culture of miniature rose[J]. Scientia Silvae Sinicae,2004,40(4):189-192.
• [46]鲁敏，汤浩茹，余世杰，雷美艳，陈小杨，丁继军.二次回归正交旋转组合设计在梨子叶不定梢再生研究中的应用[J].果树学报，2011,28(1):171-175.LU Min,TANG Haoru,YU Shijie,LEI Meiyan,CHEN Xiaoyang,DING Jijun. Application of quadratic regressive orthogonal rotational combinational design for regeneration study of pear cotyledon adventitious shoots[J]. Journal of Fruit Science,2011,28(1):171-175.
• [47] KEPENEK K,KAROGLU Z. The effects of paclobutrazol and daminozide on in vitro micropropagation of some apple(Malus domestica)cultivars and M9-rootstock[J]. African Journal of Biotechnology,2011,10(24):4851-4859.