Presnail J K, Hoy M A
Department of Entomology, University of California, Berkeley, USA.
Exp Appl Acarol. 1994 Jun;18(6):319-30. doi: 10.1007/BF00116313.
The persistence of DNA injected into two species of adult female phytoseiids and its transmission to serial eggs deposited by them was assessed by the polymerase chain reaction (PCR). The effect of DNA concentration on persistence and transmission was examined in Metaseiulus occidentalis. M. occidentalis females were microinjected with plasmid DNA at three different concentrations (250, 500, 750 ng microL-1) and allowed to deposit one to five eggs before the females and their last eggs were analyzed. Plasmid DNA was found in 82% of the females assayed and in 70% of all the eggs analyzed (including the fifth eggs produced after microinjection). Transmission of DNA to multiple eggs was also examined in Amblyseius finlandicus. Females of this species are less traumatized by microinjection allowing analysis of transmission over a more extended number of eggs. Females were microinjected and allowed to deposit eggs until their death. DNA from every fifth egg was analyzed by the PCR. PCR products were amplified from 51% of the eggs and from all egg classes except the 30th egg. The persistence and presence of plasmid DNA in both eggs and females suggests that (1) maternal microinjection is a more efficient method for DNA delivery than traditional egg microinjection, (2) it may be possible to isolate transformants from fewer maternally-microinjected females than originally expected, and (3) maternal microinjection could be useful as a DNA delivery system in other phytoseiids.
通过聚合酶链反应（PCR）评估了注入两种成年雌性植绥螨体内的DNA的持久性及其向它们所产后续卵的传递情况。在西方盲走螨中研究了DNA浓度对持久性和传递的影响。给西方盲走螨雌性个体显微注射三种不同浓度（250、500、750 ng μL-1）的质粒DNA，让它们产下1至5枚卵，然后对雌性个体及其最后产下的卵进行分析。在所检测的雌性个体中，82%发现含有质粒DNA，在所有分析的卵（包括显微注射后产下的第5枚卵）中，70%含有质粒DNA。还在芬兰钝绥螨中研究了DNA向多枚卵的传递情况。该物种的雌性个体受显微注射的创伤较小，从而可以分析更多枚卵的传递情况。对雌性个体进行显微注射，让它们产卵直至死亡。每隔5枚卵取一枚，通过PCR分析其中的DNA。从51%的卵以及除第30枚卵之外的所有卵组中扩增出了PCR产物。质粒DNA在卵和雌性个体中的持久性及存在情况表明：（1）母本显微注射是一种比传统的卵显微注射更有效的DNA导入方法；（2）从母本显微注射的雌性个体中分离转化体所需的数量可能比原先预期的要少；（3）母本显微注射作为一种DNA导入系统在其他植绥螨中可能有用。
