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首页> 外文会议>Proceedings of the 12th International Rapeseed Congress: Sustainable Development in Cruciferous Oilseed Crops Production >High frequency production of microspore derived doubled haploids (DH) and its application for developing low glucosinolate lines in Indian Brassica juncea
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High frequency production of microspore derived doubled haploids (DH) and its application for developing low glucosinolate lines in Indian Brassica juncea

机译：小孢子衍生双倍体单倍体（DH）的高频生产及其在芥菜油菜低硫代芥子油系中的应用

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摘要

The Indian varieties of Brassica juncea are high in erucic acid (EA) and glucosinolate (GSL) contents and are, therefore,recessive genes and hence, can be conveniently handled only through the production of doubled haploids. We developed an efficient protocol for the production of microspore derived doubled haploids in B. juncea, with reasonably high embryogenic and very high embryo conversion frequencies and applied it in a backcross breeding programme for the introgression of low glucosinolate trait from an un-ad apted canola quality B. juncea line Heera to a popular Indian variety Varuna. The parents differed significantly in their average embryogenic response and that of the F1DH was close to Varuna Microspores treated with colchicine at the initiation of culture for 24h showed 65-70% doubled haploid production. Maintenance of high humidity in the immediate environment of the cultured microspores was crucial for the induction of embryogenesis. Addition of activated charcoal after colchicine treatment improved the embryogenic frequency. Subjecting the fully differentiated embryos to ABA at low temperature (15°C) for physiological maturation eliminated embryo malformations such as secondary embryogenesis,The doubled haploids were transferred to the field without any prior hardening with a mortality rate of about 10%. Till date,following this protocol through five backcross generations coupled with stringent selections, we have developed several low GSL (≤10 μmol/g of seed) lines in B. juncea.
机译：印度芥菜油菜中的芥酸（EA）和芥子油苷（GSL）含量高，因此是隐性基因，因此只能通过产生双倍单倍体来方便地处理。我们开发了一种有效的方案，用于在芥菜气单胞菌中产生微孢子衍生的双倍单倍体，并具有相当高的胚发生频率和非常高的胚转化频率，并将其应用于回交育种程序中，以从不适应的双低油菜籽中渗入低芥子油苷特性。优质的芥菜线Heera到印度流行的Varuna。亲本的平均胚胎发生反应差异显着，F1DH的亲和力与秋水仙碱在培养开始24小时后所用的Varuna小孢子接近，表明单倍体产量增加了65-70％。在培养的小孢子的直接环境中维持高湿度对于诱导胚胎发生至关重要。秋水仙碱处理后添加活性炭改善了胚发生频率。将完全分化的胚胎在低温（15°C）下进行ABA进行生理成熟，消除了胚胎继发胚等畸形，双倍的单倍体被转移到田间，没有任何事先硬化，死亡率约为10％。迄今为止，在经过五代回交和严格选择之后，我们已经在芥菜油菜中开发了几种低GSL（≤10μmol/ g种子）品系。

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