Department of Biology, Washington University, 63130, St. Louis, MO, U.S.A..
Plant Mol Biol. 1987 Jan;8(1):13-22. doi: 10.1007/BF00016430.
The treatment of barley aleurone layers with gibberellic acid (GA3) results in the synthesis of two groups of α-amylase isozymes. Addition of abscisic acid (ABA) at the same time as GA3 inhibited the synthesis of both groups of isozymes. However, midcourse ABA addition (12 h or later after GA3) had a more inhibitory effect on the high pI α-amylase group than on the low pI α-amylase group. This midcourse inhibition was detectable within 2 h of ABA addition. Northern analysis results using cDNA probes for the high pI and low pI α-amylase groups paralleled the protein synthesis results for both isozyme groups. High pI α-amylase mRNA levels began to decrease within 2 h of midcourse ABA treatment and were less than 10% of the original level by 4 h. The levels of low pI α-amylase mRNA were decreased less by midcourse ABA addition than were high pI mRNA levels. Cordycepin and cycloheximide blocked the effects of midcourse ABA addition on α-amylase mRNA. These observations indicate that ABA inhibits α-amylase expression at the pretranslational level and that protein and RNA synthesis are required for midcourse ABA action to occur. Our results also show that α-amylase mRNA, which has been thought to be very stable, is degraded after midcourse ABA treatment.
用赤霉素(GA3)处理大麦糊粉层会导致两组α-淀粉酶同工酶的合成。同时添加脱落酸(ABA)会抑制这两组同工酶的合成。然而,中途添加 ABA(GA3 后 12 小时或更晚)对高 pI α-淀粉酶组的抑制作用比对低 pI α-淀粉酶组更强。这种中途抑制在 ABA 添加后 2 小时内即可检测到。使用高 pI 和低 pI α-淀粉酶组的 cDNA 探针进行的 Northern 分析结果与两种同工酶组的蛋白质合成结果一致。高 pI α-淀粉酶 mRNA 水平在中途 ABA 处理后 2 小时内开始下降,4 小时时降至原始水平的 10%以下。与高 pI mRNA 水平相比,中途添加 ABA 对低 pI α-淀粉酶 mRNA 的降低程度较小。虫草素和环己酰亚胺阻断了中途添加 ABA 对α-淀粉酶 mRNA 的影响。这些观察结果表明,ABA 在翻译前水平抑制α-淀粉酶的表达,并且需要蛋白质和 RNA 合成才能使中途 ABA 发挥作用。我们的结果还表明,α-淀粉酶 mRNA 在中途添加 ABA 后会降解,尽管之前认为它非常稳定。