Method for improving low-nitrogen tolerance character of wheat crops
A crop and wheat technology, applied in the field of crop breeding, can solve problems such as environmental pollution, economic benefit decline, and economic loss
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Embodiment 1
[0136] Embodiment 1: the low-nitrogen stress-resistant cultivation of barley microspores
[0137] "Hua 30" was used as the test material, and the tassels whose microspores were in the early and middle stages of uninucleate development were selected from Daejeon, and put in the refrigerator for 15 days. When inoculating, the tassels were sterilized with a saturated bleach solution for 15 minutes, and washed 3 to 4 times with sterile water. Each test tube received 10 tassels, poured 15ml of mannitol 60g / L, CaCl 2 1.1g / L, MES 0.976g / L extract, use a 3000rpm high-speed disperser to spin cut at a high speed, filter with a 150-mesh screen, and centrifuge the filtrate at a low speed of 800rpm, repeat 3 times, and collect microspores. The microspores were pretreated with the extract at 25°C in the dark for 2 days. Induction medium without KNO 3 and (NH 4 ) 2 SO 4 N 6 The culture medium is basic medium, which is supplemented with 2,4,5-T 1.0mg / L, KT 0.5mg / L, maltose 90g / L, gluta...
Embodiment 2
[0138] Embodiment 2: Acquisition of tolerant low nitrogen regeneration plants
[0139] After 21 days of induction culture, the embryoid bodies were transferred to the differentiation medium. Differentiation medium without KNO 3 and NH 4 NO 4 The 2 / 3MS is the basic medium, which is added with 6-BA 0.5mg / L, KT 1.5mg / L, NAA0.05mg / L and maltose 30g / L, glutamine and hydrolyzed casein each 400mg / L, pH is 5.8. Under the conditions of 25±1° C. and 10-12 hours of light per day, the differentiation stress culture is carried out for 25-30 days until green regenerated plants are differentiated.
Embodiment 3
[0140] Embodiment 3: the rooting culture of tolerance low nitrogen regeneration plant
[0141] The differentiated regenerated plants were selected and transferred to 1 / 2MS rooting and seedling growth medium with pH 5.8 supplemented with 400 mg / L of glutamine and 400 mg / L of hydrolyzed casein, 5.0 mg / L of paclobutrazol (MET) and 20 g / L of sucrose. Under the conditions of 25±1° C. and 10 to 12 hours of light per day, the cultivation of rooted and strong seedlings is carried out for 25 to 30 days.
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