Acta Agrestia Sinica ›› 2017, Vol. 25 ›› Issue (3): 582-589,603.DOI: 10.11733/j.issn.1007-0435.2017.03.019

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Study on Regeneration System Optimization of Cynodon dactylon ‘Xinnong No.1’ Using Orthogonal Array Testing

LIU Li, LI Pei-ying   

  1. College of Prataculture and Environmental Science, Xinjiang Agricultural University, Xinjiang Key Laboratory of Grassland Resources and Ecology, Urumqi, Xinjiang 830052, China
  • Received:2016-06-03 Revised:2017-03-27 Online:2017-06-15 Published:2017-08-29

应用正交试验法优化‘新农1号’狗牙根再生体系

刘莉, 李培英   

  1. 新疆农业大学草业与环境科学学院 新疆草地资源与生态自治区重点实验室, 新疆 乌鲁木齐 830052
  • 通讯作者: 李培英
  • 作者简介:刘莉(1987-),女,宁夏中卫人,硕士研究生,研究方向为草坪草育种,E-mail:540425658@qq.com
  • 基金资助:

    新疆高新技术课题(201111122);新疆草地资源与生态实验室开放课题(XJDX209-2013-1);新疆科技厅重大专项(201230116-4)资助

Abstract:

With the mature seeds of Cynodon dactylon ‘Xinnong No.1’ as materials, the effects of the hormones and additives on callus induction, callus subculture and callus differentiation were investigated through orthogonal design method and the regeneration system was optimized. For callus induction of ‘Xinnong No.1’, the influence sequence of three factors was 2,4-D > 6-BA > Proline. MS medium supplemented with 2.0 mg·L-1 2,4-D+0. 01 mg·L+1 6-BA+200 mg·L-1 Proline was the optimum induction medium and callus induction rate could reach 69.5%. During callus subculture, the influence sequence of three factors for induction rate of embryonic callus, callus proliferation rate and browning rate were different. The influence sequence for induction rate of embryonic callus, callus proliferation rate and browning rate was 2,4-D > ABA > 6-BA, 6-BA > ABA > 2,4-D, and 2,4-D > 6-BA > ABA, respectively. Comprehensive analysis showed that MS medium supplemented with 2.0 mg·L-12,4-D+0.2 mg·L-1 6-BA+2.0 mg·L-1 Proline was the optimized subculture medium. The embryogenic callus rate and callus proliferation rate were 71.67% and 61.67%, respectively, and the browning rate was low. Result of bud differentiation showed that MS medium supplemented with 1.0 mg·L-16-BA was the optimized differentiation medium and plantlet formation strength could reach 49.44%.

Key words: Cynodon dactylon ‘Xinnong No.1’, Orthogonal experimental design, Regeneration system

摘要:

以‘新农1号’狗牙根(Cynodon dactylon(L.) Pers.)成熟颖果为材料,通过正交法探讨激素、外源添加物等对狗牙根愈伤诱导、继代及分化的影响,优化‘新农1号’狗牙根再生体系。结果表明:各因子对‘新农1号’狗牙根愈伤诱导的影响程度为2,4-D > 6-BA > 脯氨酸;优化诱导培养基为添加2,4-D(2.0 mg·L-1)+ 6-BA(0.01 mg·L-1)+ 脯氨酸(200 mg·L-1)的MS培养基,此时愈伤诱导率达69.5%;对继代增殖而言,各因子对胚性愈伤诱导率、愈伤增殖率、褐化率的影响程度不同,对胚性愈伤诱导率的影响程度为2,4-D > ABA > 6-BA,对愈伤增殖率的影响程度为6-BA > ABA > 2,4-D,对褐化率的影响程度为2,4-D > 6-BA > ABA。综合3个指标结果认为,‘新农1号’狗牙根愈伤继代的优化培养基为2,4-D(2.0 mg·L-1) +6-BA(0.20 mg·L-1)+ ABA(2.0 mg·L-1)的MS培养基,其胚性愈伤率和愈伤增殖率分别可达71.67%和61.67%,且褐化率较低;对分化而言,优化的分化培养基为MS+6-BA(1.0 mg·L-1),小植株形成强度可达49.44%。

关键词: ‘新农1号’狗牙根, 正交试验法, 再生体系

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