Orthogonal array design for optimization of an artificial medium for in vitro rearing of Trichogramma dendrolimi
Orthogonal array design for optimization of an artificial medium for in vitro rearing of Trichogramma dendrolimi
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正交阵列设计优化松毛虫赤眼蜂体外培养人工培养基
DOI：
10.1111/eea.12194
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发表时间：
2014-07-01
影响因子：
1.9
通讯作者：
Li, Liying
中科院分区：
文献类型：
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作者：
Lu, Xin;Han, Shichou;Li, Liying
赤眼蜂在人工培养基上的成功发育还与寄主昆虫衍生成分以外的成分的存在和比例有关。本研究以松毛虫赤眼蜂Trichogramma endrolimi MatSumura(膜翅目：赤眼蜂科)为材料，采用三水平六因素三水平正交表，对松毛虫赤眼蜂的寄生、幼虫发育、化蛹和成虫羽化等参数进行监测，以确定人工培养基组分的最重要成分，并改进松毛虫赤眼蜂体外大量繁殖的培养基料。结果表明，人工培养基中的6种成分对杜鹃幼虫的生物学参数有不同程度的影响，分别为天蚕(Antheraea pernyi(Guerin-Meneville)(鳞翅目：土蛾科))、蛋黄、10%麦芽乳液、尼森海默盐液、海藻糖和灭菌水。统计分析表明，海藻糖和Neisenheimer‘s盐溶液、10%麦芽乳液和珀氏小蜂蛹血淋巴是人工培养液的主要成分，以寄生率和化蛹率、每个人工卵中发育的幼虫数量、成虫羽化率和正常成虫数量为指标。后续的生物测定和优化配方的选择证实了正交表分析预测的优化方法的有效性，表明该方法在食虫性昆虫人工饲料的选择中是有效的。在优化的人工培养基上，每张卵卡(每张含20枚人工卵)平均成虫羽化率和正常成虫数分别为88.8%和956只雌蜂。后一种培养基比以前开发的任何一种培养基都要好，因此有可能在体外大规模繁殖松毛虫。
Successful complete development of Trichogramma species on artificial media is also related to the presence and proportions of ingredients other than host insect-derived components. In this study, an orthogonal array with six factors at three levels was performed and parameters of parasitism, larval development, pupation, and adult emergence were monitored to reveal the most important components of an artificial medium and to improve the medium for in vitro mass rearing of Trichogramma dendrolimi Matsumura (Hymenoptera: Trichogrammatidae). Results indicated that biological parameters of T.dendrolimi were affected differently by six ingredients of the artificial medium: pupal hemolymph of Antheraea pernyi (Guerin-Meneville) (Lepidoptera: Saturniidae), egg yolk, 10% malted milk solution, Neisenheimer's salt solution, trehalose, and sterile water. Statistical analysis indicated that trehalose and Neisenheimer's salt solution, 10% malted milk solution, and pupal hemolymph of A.pernyi were the main ingredients of the artificial medium based on rates of parasitism and pupation, the number of larvae developing in each artificial egg, adult emergence rate, and the number of normal adults produced. A follow-up bioassay with a selection of optimized formulas confirmed the validity of the optimization as predicted by the orthogonal array analysis, indicating the usefulness of this method for selecting artificial diets for entomophagous insects. Adult emergence rate of the parasitoid and total number of normal adults produced per egg card (each containing 20 artificial eggs) averaged 88.8% and 956 females on the best performing optimized artificial medium, consisting of 3ml pupal hemolymph of A.pernyi, 2.5ml egg yolk, 1ml 10% malted milk solution, 1ml Neisenheimer's salt solution, 0.1g trehalose, and 1.5ml sterile water. The latter medium was superior to any formerly developed medium and may thus have potential for the in vitro mass rearing of T.dendrolimi.