CSIRO, Division of Plant Industry, 2601, Canberra, ACT, Australia.
Theor Appl Genet. 1993 Dec;87(4):455-63. doi: 10.1007/BF00215091.
This paper reports on the production of intergeneric somatic hybrid plants between two sexually incompatible legume species. Medicago sativa (alfalfa, lucerne) leaf protoplasts were inactivated by lethal doses of iodoacetamide. Onobrychis viciifolia (sainfoin) suspension-cell protoplasts were gamma-irradiated at lethal doses. Following electrofusion under optimized conditions about 50,000 viable heterokaryons were produced in each test. The fusion products were cultured with the help of alfalfa nurse protoplasts. Functional complementation permitted only the heterokaryons to survive. A total of 706 putative heterokaryon-derived plantlets were regenerated and 570 survived transplantation to soil. Experimentation was aimed at the introduction of proanthocyanidins (condensed tannins) from sainfoin, a bloat-safe plant, to alfalfa, a bloat-causing forage crop; however, no tannin-positive regenerant plants were detected. Most regenerant plants have shown morphological differences from the fusion parents, although, as expected, all resembled the "recipient" parent, alfalfa. Southern analysis using an improved total-genomic probing technique has shown low levels of sainfoin-specific DNA in 43 out of 158 tested regenerants. Cytogenetic analysis of these asymmetric hybrids has confirmed the existence of euploid (2n=32; 17%) as well as aneuploid (2n=30, 33-78; 83%) plants. Pollen germination tests have indicated that the majority of the hybrids were fertile, while 35% had either reduced fertility or were completely sterile.
本文报告了两种性不亲和豆科植物之间产生属间体细胞杂种植物的情况。通过致死剂量的碘乙酰胺使紫花苜蓿（紫花苜蓿，三叶草）叶原生质体失活。用致死剂量的γ射线辐照红豆草（苦马豆）悬浮细胞原生质体。在优化条件下电融合后，每个测试中产生约 50,000 个活异核体。在紫花苜蓿保姆原生质体的帮助下培养融合产物。功能互补仅允许异核体存活。总共产生了 706 个假定的异核体衍生的胚状体，并存活了 570 个移植到土壤中。实验旨在将来自不易膨胀的安全植物苦马豆的原花青素（缩合单宁）引入到易膨胀的饲料作物紫花苜蓿中；然而，没有检测到单宁阳性再生植物。尽管大多数再生植物与融合亲本在形态上存在差异，但正如预期的那样，它们都与“受体”亲本紫花苜蓿相似。使用改进的全基因组探针技术进行的Southern 分析表明，在 158 个测试的再生植物中有 43 个显示出苦马豆特异性 DNA 的低水平。这些不对称杂种的细胞遗传学分析证实了存在整倍体（2n=32；17%）和非整倍体（2n=30，33-78；83%）植物。花粉萌发试验表明，大多数杂种是可育的，而 35%的杂种要么可育性降低，要么完全不育。
