Effect of Cultivar, Incubation Temperature, and Stage of Microspore Development on Anther Culture in Wheat and Triticale

Abstract

Effects of incubation temperature and developmental stage of microspores on polyhaploid production in three wheat cultivars 'Pavon 76', 'Kitt', and 'Chris' and one tnticale cultivar, 'T8r, were studied using a one-step medmm. Calh failing to differentiate on the one-step medium were placed on a medium containing 1 mg/1 indole-3-acetic acid (IAA) and 2 mg/I 6-furfurylaminopurine (KIN). Anthers containing either early-or late-uninucleate microspores were incubated in dark at 26, 28 or 32 °C for 3 days prior to transfer to 26 °C. Averaged over temperatures and microspore stages, frequency of calli and green plantiets were 8.9 % and 3.4 %, respectively, for wheat cultivar 'Pavon 76', 8.4% and 1.6% for cultivar 'Kitt', 4.5 % and 0.25 % for cultivar 'Chris', and 2.9 % and 0.12 % for the triticale cultivar 'T8r. However, cultivar-by-developmentalstage interaction was significant for frequency of callus induction. Temperature had no significant effects on callus induction and plantlet regeneration. Anthers containing early-uninucleate microspores produced no polyhaploids. Key words: Triticum aestivum -anther cultureculture conditions -temperature response -stage of microspores. Contribution No. 89-283-J from the Kansas Agricultural Experiment Station; Kansas State University, Manhattan.Among the conditions and variables used in an anther culture system, incubation temperature and stage of microspore development are important factors. Since wheat requires cool conditions for germination and initial growth, early research on anther culture generally was carried out at relatively low temperatures (20 to 27 °C) (OuYANG et al. 1973). Culture temperature for inducing microspore callus in wheat ranges from 26 to 30 °G, but 28 to 30 °C was reported best for most cultivars (OUYANG et al. 1983). The responses of anther culture to culture temperature were also affected by growth conditions of anther-donor plants; highest yield of green pollen piantlets appeared at a culture temperature of 30 ^G when the anther-donor plants had been grown in the greenhouse, but was at 32 °G when the donor plants were grown in field (OUYANG 1986, OUYANG et al. 1987. Two wheat cultivars produced 40 % more microspore calli and green plantiets at 30 than at 25 °C (HUANG 1987). High yields of wheat pollen calli and piantlets were obtained at 2% to 32 °G for the cultivar 'Ghris' (Ho et al. 1978). For spring wheat, callus yield was increased when anthers were incubated at 32 °C for 6 days prior to incubation at 26 °G (LI et al. 1988). Variation in intensity and duration of low-temperature U.S. Copyright Clearance Center Code Statement: 0 1 79-954 1 /90/0504-0332$02.50/0

Effect of Sugars in Wheat Anther Culture Media

“…The genotype and I-sucrose results for embryoid initiation are similar to YUAN et al (1990), SIMONSON and BAENZIGER (1992), and RYBCZYNSKI et al (1991) indicating the repeatability of our anther culture system. The differential response of genotypes to wheat anther culture media I-sugars was observed with other cultivars and media for sucrose , HASSAWI and LIANG 1990, HASSAWI et al 1990, BALL et al 1992) for sucrose and maltose (ZHOU et al 1991), for maltose (ORSHINSKY et al 1990), and for maltose, fructose, and glucose (LAST and BRETTELL 1990).…”

“…'Pavon 76' and 'Centurk', both known to be responsive genotypes, produced a similar number of green plants per embryoid (0.44 and 0.38, respectively) while 'Arapahoe' produced only 0.02 green plants per embryoid ( Table 2). The frequency of plant regeneration in wheat is well known to be genotype dependent , LIANG and MCHUGEN 1987, HASSAWI et al 1990, YUAN et al 1990, HASSAWI and LIANG 1991b. BALL et al 1992).…”

“…'Pavon 76' produced more albinos per embryoid (0.09) than 'Centurk' (0.04). The number of albino plants is known to be affected by genotype (ANDERSEN et al 1987, HASSAWI et al 1990, OUYANG et al 1983). The I-sugars (excluding fructose) with R-sucrose did not differ for albino plants per embryoid (Table 2).…”

“…The ability of wheat embryoids to regenerate plants is influenced by I-sugars (ORSHINSKY et al 1990, ZHOU et al 1991, BALL et al 1992), growth regulators (HASSAWI et al 1990), and amino acid supplementation (CHU and HILL 1988). I-sugar type and concentration in initiation media also partially determined the ratio of green : albino plants (KUHLMANN and FOROUGHI-WEHR 1989, LAST and BRETTELL 1990, ZHOU et al 1991 Regeneration media usually have the same composition as initiation media except for lower concentrations of growth regulators and sugars.…”

Isolated microspore culture of Canadian 6× triticale cultivars

“…Cold pretreatment at 4°C for 1-3 weeks alone or in combination with heat shock (+32°C for 3 days) or mannitol (175.6 mM) induced starvation and a prolonged cold stress of 3 weeks improved androgenetic response (Immonen and Robinson, 2000). However, Hassawi and Liang (1990) reported that heat shock treatment right after harvesting had no significant effect on triticale callus induction and plantlet regeneration. The culture media used for anther culture have been 190-2, N6, MN6, B5, K1, KM, Potato II medium and C17 (Ryoppy, 1996).…”