Laboratory of Plant Cell and Molecular Biology, Department of Vegetable Crops, University of Florida, 32611, Gainesville, FL, USA.
Theor Appl Genet. 1990 May;79(5):609-17. doi: 10.1007/BF00226873.
Immature embryos, inflorescences, and anthers of eight commercial cultivars of Triticum aestivum (wheat) formed embryogenic callus on a variety of media. Immature embryos (1.0-1.5 mm long) were found to be most suitable for embryogenic callus formation while anthers responded poorly; inflorescences gave intermediate values. Immature embryos of various cultivars showed significant differences in callus formation in response to 11 of the 12 media tested. No significant differences were observed when the embryos were cultred under similar conditions on MS medium with twice the concentration of inorganic salts, supplemented with 2,4-D, casein hydrolysate and glutamine. Furthermore, with inflorescences also no significant differences were observed. Explants on callus formation media formed two types of embryogenic calli: an off-white, compact, and nodular callus and a white compact callus. Upon successive subcultures (approximately 5 months), the nodular embryogenic callus became more prominent and was identified as 'aged callus'. The aged callus upon further subculture, formed an off-white, soft, and friable embryogenic callus. Both the aged and friable calli maintained their embryogenic capacity over many subculture passages (to date up to 19 months). All embryogenic calli (1 month old) from the different callus-forming media, irrespective of expiant source, formed only green shoots on regeneration media that developed to maturity in the greenhouse. There were no significant differences in the response of calli derived from embryos and inflorescences cultured on the different initiation media. Also, the shoot-forming capacity of the cultivars was not significantly different. Anther-derived calli formed the least shoots. Aged and friable calli on regeneration media also formed green shoots but at lower frequencies. Plants from long-term culture have also been grown to maturity in soil.
八种商业化小麦(Triticum aestivum)品种的未成熟胚胎、花序和花药在多种培养基上形成了胚性愈伤组织。研究发现,1.0-1.5 毫米长的未成熟胚胎最适合形成胚性愈伤组织,而花药反应较差;花序居中。在 12 种测试培养基中,11 种培养基中不同品种的未成熟胚胎在愈伤组织形成方面表现出显著差异。在 MS 培养基上,以两倍浓度的无机盐、添加 2,4-D、水解酪蛋白和谷氨酰胺,在相似条件下培养胚胎时,未观察到显著差异。此外,花序也未观察到显著差异。在愈伤组织形成培养基上的外植体形成了两种类型的胚性愈伤组织:一种是灰白色、紧实、结节状的愈伤组织和一种是白色紧实的愈伤组织。经过连续继代培养(约 5 个月),结节状胚性愈伤组织变得更加突出,并被鉴定为“老化愈伤组织”。进一步继代培养后,老化的愈伤组织形成了灰白色、柔软、易碎的胚性愈伤组织。老化和易碎的愈伤组织在多次继代培养中(迄今为止多达 19 个月)保持其胚性能力。所有来自不同愈伤组织形成培养基的胚性愈伤组织(1 个月龄),无论外植体来源如何,在再生培养基上仅形成绿芽,这些绿芽在温室中发育成熟。在不同起始培养基上培养的胚胎和花序来源的愈伤组织的响应没有显著差异。此外,品种的芽形成能力也没有显著差异。花药来源的愈伤组织形成的芽最少。再生培养基上的老化和易碎的愈伤组织也形成绿芽,但频率较低。长期培养的植株也在土壤中生长成熟。