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秋季采摘柑橘成熟果实并剥取种子,将小粒种籽和瘪籽播于MT培养基,待萌发长出2～3片真叶时,取其叶片检测倍性;将饱满种籽播种于露地,对其幼苗首先通过形态筛选获得似多倍体的植株,然后取其叶片用流式细胞仪进行倍性检测;最后对获得的多倍体植株进行SSR分子鉴定。试验结果表明,获得早金甜橙、卡特夏橙、塔罗科血橙、日辉橘、Ortanique橘橙、默科特橘橙、华农本地早橘、朱红橘和Flame葡萄柚等9个品种的四倍体分别为6、1、3、12、3、6、2、1和10株;获得早金甜橙、塔罗科血橙、Itabori甜橙和Ortanique橘橙等4个品种的三倍体分别为1、1、2、4株。用13对多态性SSR引物对胚抢救获得的三倍体和四倍体的来源进行鉴定,发现早金甜橙、Itabori甜橙的三倍体再生植株为异源三倍体;塔罗科血橙、Ortanique橘橙的三倍体可能为同源三倍体;早金甜橙和Ortanique橘橙的四倍体可能为同源四倍体;塔罗科血橙四倍体为异源四倍体。采用37对多态性引物对从饱满种籽中获得的8个品种四倍体的来源进行鉴定,发现均为同源四倍体。这些自然发掘的多倍体资源对柑橘无核育种和相关基础研究具有重要价值。
Mature citrus fruits are harvested in autumn and the seeds are stripped. Small seeds and shriveled seeds are seeded on MT medium. When 2-3 true leaves are to be sprouted, ploidy is taken from the leaves of the leaves. The full seeds are planted in the open field, First of all, the seedlings were seeded with polyploid plants through morphological screening. The leaves were then analyzed by flow cytometry for ploidy detection. Finally, the polyploid plants were identified by SSR. The results showed that 9 varieties of early orange, Carter orange, Taro blood orange, Rihui orange, Ortanique orange, Merck orange, Huarong native orange, Vermilion and Flame grapefruit were obtained Tetraploids were 6, 1, 3, 12, 3, 6, 2, 1 and 10 strains respectively; obtained three golds of early gold sweet orange, taro blood orange, Itabori sweet orange and Ortanique tangerine orange Body were 1, 1, 2, 4 strains. Thirteen pairs of polymorphic SSR primers were used to identify the origins of the triploid and tetraploid embryos. It was found that the triploid plants of early golden orange and Itabori sweet orange were allotriploid; Blood oranges, Ortanique orange orange triploids may be autotetraploid; early gold orange and Ortanique orange orange tetraploid may be autotetraploid; Tarot blood orange tetraploid heterotetra Ploidy. 37 pairs of polymorphic primers were used to identify the origins of tetraploids of eight cultivars obtained from full seeds and found to be all autotetraploid. These naturally-found polyploid resources are of great value to nuclear citrus seedless breeding and related basic research.