Development of an Asymbiotic Reference Line for Pea cv. Bohatýr by De Novo Mutagenesis
Development of an Asymbiotic Reference Line for Pea cv. Bohatýr by De Novo Mutagenesis
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通过 De Novo Mutagenesis 开发豌豆品种 Bohatýr 的非共生参考品系
DOI：
10.2135/cropsci2004.0670
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发表时间：
2005
期刊：
影响因子：
2.3
通讯作者：
J. Vondrys
中科院分区：
文献类型：
--
作者：
K. Novák;Martin Slajs;E. Biedermannová;J. Vondrys
非共生和共生品系对的比较（差异技术）是确定共生固氮对豆科作物产量贡献的可靠和可行的方法。为获得商品豌豆（PisumsativumL.）用CV启动诱变程序。博哈蒂尔在封闭系统中优化了甲基磺酸乙酯（EMS）处理大种子的工艺，以扩大工艺规模并提高安全性。非预浸种子暴露于0.15%（w/w）EMS溶液中18小时，导致30%的田间出苗在M1代和1.04%的叶绿素突变体和3.64%的突变体分类为共生的M2。然而，只有48.4%的生长室检测和21.6%的现场检测推定共生突变体被发现是稳定的遗传变异的M3至M5。22个初步表征的稳定突变体包括一个nonacetulating，一个减少acetulation，一个显着延迟acetulation，和19个受影响的结节发育和功能的线。在受控条件下和田间试验中的进一步评价确定了非固定线Budfix 2作为共生固氮的田间估计的合适参考线。当在非共生条件下生长时，该品系显示出生理上等同于原始栽培品种，即，当共生断层被外源矿质氮补偿时。Budfix 2的可用性允许在特定豌豆栽培品种Bohatyr中以及在性状渐渗后在其他品种中直接应用差异方法。在寻找参考系的过程中产生的额外的共生突变体预计将增强可用于根瘤菌形成的遗传解剖的变异性。
Comparison of asymbiotic and symbiotic pairs of lines (difference technique) is a reliable and accessible method for determining the contribution of symbiotic nitrogen fixation to the yield of legume crops. To obtain an isogenic reference line for a commercial pea (Pisum sativum L.) a mutagenesis program was initiated with cv. Bohatyr. Ethyl methanesulfonate (EMS) treatment of large seeds was optimized in a closed system to scale-up the process and to improve safety. The 18-h exposure of non-presoaked seeds to 0.15% (w/w) EMS solution resulted in 30% field emergence in the M 1 generation and in 1.04% of chlorophyll mutants and 3.64% of mutants classified as symbiotic in the M 2 . However, only 48.4% of the growth chamber-detected and 21.6% of the field-detected presumable symbiotic mutants were found to be stable genetic deviations in the M 3 to M 5 . Twenty-two preliminary characterized stable mutants comprised one nonnodulating, one with reduced nodulation, one with markedly delayed nodulation, and 19 lines with affected nodule development and function. Further evaluation under controlled conditions and in field trials identified the nonfixing line Budfix2 as a suitable reference line for field estimates of symbiotic nitrogen fixation. This line was shown to be physiologically equivalent to the original cultivar when grown under asymbiotic conditions, i.e., when the symbiotic fault was compensated for by exogenous mineral nitrogen. The availability of Budfix2 allows for direct application of the difference method in the particular pea cultivar Bohatyr, as well as in further varieties after trait introgression. The additional symbiotic mutants generated in the course of the search for a reference line are expected to enhance the variability available for the genetic dissection of rhizobial nodulation.