【目的】 明确斜纹夜蛾Spodoptera litura交配次数及交配顺序对父权分配的影响。【方法】 利用微卫星标记对不同多次交配处理后的斜纹夜蛾子代进行亲权鉴定。实验筛选出5对微卫星标记，共采集271头斜纹夜蛾样本，其中母本21头，父本40头，子代210头。【结果】 微卫星标记的等位基因数、期望杂合度、多态信息含量的平均值分别为4.800、0.647、0.596。鉴定置信度均高于80%，其中106头的鉴定置信度超过95%。斜纹夜蛾雌虫与2头雄虫分别交配1次，后交配雄虫的子代父权比例为78%；雌虫与1头雄虫交配2次，再与另一头雄虫交配1次，2头雄虫的子代父权比例均为50%；雌虫与1头雄虫交配1次，再与另一头雄虫交配2次，后交配雄虫的子代父权比例高达90%；雌虫与3头不同雄虫分别交配1次，3头雄虫子代父权比例分别为5%、20%和75%。【结论】 斜纹夜蛾交配次数和交配顺序均会影响父权比例。交配次数可以增加子代的父权比例，后交配的雄性拥有更高的父权比例。

[Objectives]  To investigate the effect of mating frequency and male mating order on the paternity of Spodoptera litura, an important global agriculture and forestry pest. [Methods]  Paternity was determined using five pairs of microsatellite markers. 271 samples were collected, including 21 from female parents, 40 from male parents and 210 from offspring. [Results]  The average number of alleles, expected heterozygosity and polymorphism content information were 4.800, 0.647 and 0.596, respectively. Confidence in estimated paternity was overall > 80%; > 95% for 106 offspring. If a female mated once with two males, 78% of the resultant offspring were the progeny of the last male to mate. If a female mated with a male twice with a male, then with another male once, 50% of the offspring were the progeny of each male. If a female mated with a male once, then with another male twice, 90% of the offspring were the progeny of the last male to mate. If a female mated once with three different males, 5%, 20% and 75% of the offspring were the progeny of the first, second and last males to mate, respectively. [Conclusion]  Both the frequency and order of mating affect paternity in S. litura. Males that mate more than once, and those that mate last, have more progeny than those that only mate once or that aren’t the last to mate.