响应面法优化欧洲冬青‘Ferox Argentea’增殖培养基大量元素配方  

Optimization of Macro-elements Composition in the Medium for in vitro Propagation of Ilex aquifolium‘Ferox Argentea’by Response Surface Methodology

作　　者：蔡正禹 文书生 田如男[1] Cai Zhengyu;Wen Shusheng;Tian Runan(Nanjing Forestry University,Nanjing 210000,P.R.China)

机构地区：[1]南京林业大学,南京210000

出　　处：《东北林业大学学报》2023年第6期53-63,共11页Journal of Northeast Forestry University

基　　金：国家林业局引进国际先进林业科学技术项目(2012-4-33);江苏高校优势学科建设工程项目(PAPD)。

摘　　要：利用响应面法探究了培养基大量元素组分和配比对‘Ferox Argentea’试管苗增殖的影响,并构建了该品种专用的增殖培养基,采用Design-Expert响应面最佳优化试验设计,以WPM培养基中5种大量元素无机盐K_(2)SO_(4)、MgSO_(4)·7H_(2)O、KH_(2)PO_(4)、NH_(4)NO_(3)、Ca(NO_(3))_(2)·4H_(2)O为自变量,接种50 d后的增殖率、株高、芽质量等级为因变量,建立数学模型分析并预测大量元素无机盐最佳优化配方。通过单因子效应分析表明,大量元素无机盐中NH_(4)NO_(3)对欧洲冬青‘Ferox Argentea’增殖率影响显著,对株高和芽质量影响极显著,Ca(NO_(3))_(2)·4H_(2)O对株高影响显著,对芽质量影响极显著,K_(2)SO_(4)、MgSO_(4)·7H_(2)O、KH_(2)PO_(4)对各指标影响均不显著。交互效应分析表明,K_(2)SO_(4)和KH_(2)PO_(4)、K_(2)SO_(4)和Ca(NO_(3))_(2)·4H_(2)O的交互作用对增殖率具有显著影响;KH_(2)PO_(4)和Ca(NO_(3))_(2)·4H_(2)O的交互作用对株高具有显著影响;K_(2)SO_(4)和MgSO_(4)·7H_(2)O、K_(2)SO_(4)和NH_(4)NO_(3)、KH_(2)PO_(4)和Ca(NO_(3))_(2)·4H_(2)O的交互作用对芽质量具有显著影响。因此,WPM培养基的大量元素无机盐含量影响欧洲冬青‘Ferox Argentea’的增殖率、株高、芽质量等级,NH_(4)NO_(3)与Ca(NO_(3))_(2)·4H_(2)O是主要影响单因素。响应面软件预测的大量元素无机盐优化配方为:K_(2)SO_(4)495.00 mg·L^(-1)、NH_(4)NO_(3)1200.00 mg·L^(-1)、MgSO_(4)·7H_(2)O 529.10 mg·L^(-1)、KH_(2)PO_(4)362.27 mg·L^(-1)与Ca(NO_(3))_(2)·4H_(2)O 1279.92 mg·L^(-1),进一步对该优化配方进行试验验证,发现验证组试管苗增殖率2.91,株高3.14 cm,芽质量等级2.76,与理论预测无显著差异,且显著优于对照组(1.76、2.44 cm、1.78)。综上,基于响应面法构建了欧洲冬青‘Ferox Argentea’专用的增殖培养基,研究结果不仅解决了该品种的试管苗增殖困难问题,也为其他冬青属植物增殖培养体系构建提供了重要的技术参考。Response Surface Methodology(RSM)was used to explore the effects of the composition and proportion of macro-elements in the medium on the proliferation of‘Ferox Argentea’test tube seedlings,and a special proliferation medium was constructed for this variety,in order to provide technical support for improving its in vitro rapid propagation efficiency and solving the propagation difficulties.The design expert response surface optimization design was adopted.With five kinds of macro-element inorganic salts K_(2)SO_(4),MgSO_(4)·7H_(2)O,KH_(2)PO_(4),NH_(4)NO_(3) and Ca(NO_(3))_(2)·4H_(2)O in WPM medium as independent variables,and the proliferation rate,plant height,and bud quality grade at 50 d after inoculation as dependent variables,a mathematical model was established to analyze and predict the optimal macro-element composition.By single factor effect analysis,NH_(4)NO_(3) among macro-element inorganic salts had a significant impact on the multiplication rate of Ilex aquifolium‘Ferox Argentea’,and had a very significant effect on plant height and bud quality.Ca(NO_(3))_(2)·4H_(2)O had a significant effect on plant height,and had a very significant effect on bud quality.K_(2)SO_(4),MgSO_(4)·7H_(2)O,and KH_(2)PO_(4) had no significant effect on each index.By the interaction analysis,the interaction of K_(2)SO_(4) and KH_(2)PO_(4),K_(2)SO_(4) and Ca(NO_(3))_(2)·4H_(2)O had a significant effect on the proliferation rate.The interaction of KH_(2)PO_(4) and Ca(NO_(3))_(2)·4H_(2)O has a significant effect on plant height.The interaction of K_(2)SO_(4) and MgSO_(4)·7H_(2)O,K_(2)SO_(4) and NH_(4)NO_(3),KH_(2)PO_(4) and Ca(NO_(3))_(2)·4H_(2)O had significant effects on shoot quality.Therefore,the macro-element composition in WPM medium can affect the proliferation rate,plant height and bud quality of I.aquifolium‘Ferox Argentea’,and NH_(4)NO_(3) and Ca(NO_(3))_(2)·4H_(2)O are the main factors.The optimized formulation of macro-element composition predicted by response surface software is K_(2)SO_(4)495.

关 键 词：欧洲冬青‘Ferox Argentea’ 增殖培养基 组织培养 大量元素 响应面法 配方优化 

分 类 号：S687[农业科学—观赏园艺]