Transgene Expression in Spruce and Poplar: From the Lab to the Field
Transgene Expression in Spruce and Poplar: From the Lab to the Field
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云杉和杨树中的转基因表达：从实验室到田间
DOI：
10.1007/978-94-011-3983-0_22
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发表时间：
1996
期刊：
影响因子：
--
通讯作者：
B. Mccown
中科院分区：
文献类型：
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作者：
D. Ellis;J. Rintamaki;K. Francis;Karl W. Kleiner;K. Raffa;B. Mccown
随着转化系统在一年生作物上的发展，出现了一个令人不安的主题：转基因表达存在极大的差异，特别是在同一培养皿中，来自不同转化事件的植物之间。如果转基因表达的这种变化发生在一年生作物上，那么长寿的多年生作物，如树木，可能会发生什么变化？为了回答这个问题，我们在实验室和田间研究了由增强的CaMV 35S启动子控制的uidA基因在两个杨树杂交种和几个转化品系的转化体中的表达。基于葡萄糖醛酸苷酶(GUS)活性的MUG分析，有一个总的趋势：GUS活性在体外最高，变异最小，而在田间种植的转基因植株中GUS活性变化最大。对两个生长季节三个不同发育阶段叶片中GUS活性的详细分析表明，uidA在杂交杨NC5339(P.albaxP.在较老的充分展开的叶片中，GUS活性在季节和年际之间相对稳定。相比之下，另一种杂交杨NM6(P.nigraxP.相对于幼叶而言，大麦的GUS活性较高。在云杉中，针叶中的GUS活性仅在针叶伸长期间检测到，但茎中的活动在整个季节都持续存在。在田间所有转基因植物中，GUS活性在初春破芽后最高，随后活性迅速下降。在云杉和杨树的有限数量的转基因株系中已经注意到fuidA的表达受到抑制。经过三年的研究，uidA的表达在任何被抑制的转基因株系中都没有恢复。使用伤害诱导启动子(PinII)控制uidA基因，证据表明转基因在杨树中的表达年复一年的稳定性并不是CaMV 35S所特有的。有趣的是，一个物种内连续两个季节的GUS活性总体水平是相似的，尽管存在季节性变化，但在木本植物多年生植物中，转基因表达的可预测的年度趋势可能是可能的。
As the development of transformation systems progresses in annual crop plants, one troubling theme has emerged: there is extreme variation in transgene expression, particularly between plants derived from individual transformation events even within the same petri dish. If such variation in transgene expression occurs in annual crops, what variation can be expected in long-lived perennial crops, such as trees? To answer this question, we have studied the expression over time, both in the lab and in the field, of theuidAgene controlled by an enhanced CaMV 35S promoter in transformants from twoPopulushybrids and several transformed lines ofPicea glauca. Based on MUG assays for ß-glucuronidase (GUS) activity, there is a general trend: the highest and least variable GUS activity occurs in vitro and the most variable GUS activity is in field grown transgenics. Detailed analysis of GUS activity in three different developmental stages of leaves during two growing seasons indicate thatuidAexpression in young leaves of hybrid poplar NC5339 (P. albaxP. grandidentata) was highly variable and that in older fully expanded leaves, GUS activity was relatively stable during the season and from year to year. In contrast, older leaves in another hybrid poplar, NM6 (P. nigraxP. maximowiczii) had higher GUS activity relative to the younger leaves. In spruce, GUS activity in needles was only detected during needle elongation, yet activity in the stems persists throughout the season. In all transgenics in the field, GUS activity was highest early in the spring just after bud break, then activity rapidly dropped. The suppression ofuidAexpression has been noted in a limited number of transgenic lines in both spruce and poplar. After three years in the field,uidAexpression has not been restored in any transgenic lines that haduidAexpression suppressed. Using a wound-inducible promoter (pinII) to control theuidAgene, evidence suggests that year-to-year stability of transgene expression in poplar is not unique to CaMV 35S. It is interesting to note that the overall level of GUS activity in two consecutive seasons was similar within a species and that despite seasonal variability, predictable yearly trends in transgene expression may be possible in woody perennials.