Identification of mutants in phosphorus metabolism
Identification of mutants in phosphorus metabolism
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DOI：
10.1111/j.1744-7348.2001.tb00091.x
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发表时间：
2001-01-01
影响因子：
2.6
通讯作者：
Raines, CA
中科院分区：
文献类型：
--
作者：
Lloyd, JC;Zakhleniuk, OV;Raines, CA
磷的有效性往往限制植物的生长。然而，对植物磷吸收和分配的调控途径和机制知之甚少。我们已经开发了一个筛选的基础上分泌的根酸性磷酸酶活性的诱导低磷胁迫，以确定突变体的拟南芥磷代谢缺陷。用肉眼检测到A.在低磷培养基上，以显色底物5-溴-4-氯-3-吲哚基磷酸盐（BCIP）为显色底物，对拟南芥幼苗进行了研究。在低磷胁迫条件下，野生型植物的根染成蓝色，因为诱导的根酸性磷酸酶裂解BLIP释放有色产物。在这些条件下，潜在的突变体被鉴定为具有白色或颈蓝色根。在筛选的约79 000株T-DNA诱变幼苗中，进一步鉴定了两株酸性磷酸酶染色减少的突变体。与野生型A相比，两者都表现出生长减少和P含量的差异。thaliana.具有最严重表型的突变体pho3在叶片内以独特的视觉模式积累了高水平的花青素和淀粉。这些突变体的表型不同于先前鉴定的两个磷突变体（pho1和pho2）和A. thaliana.这表明，筛选方法是强大的，并可能导致进一步的突变体的鉴定，增加我们对磷营养的理解的潜力。
Phosphorus availability is often limiting for plant growth. However, little is known of the pathways and mechanisms that regulate phosphorus (P) uptake and distribution in plants. We have developed a screen based on the induction of secreted root acid phosphatase activity by low-P stress to identify mutants of Arabidopsis thaliana with defects in P metabolism. Acid phosphatase activity was detected visually in the roots of A. thaliana seedlings grown in vitro on low-P medium, using the chromogenic substrate, 5-bromo-4-chloro-3-indolyl-phosphate (BCIP). In low-P stress conditions the roots of wildtype plants stained blue, as the induced root acid phosphatase cleaved BLIP to release the coloured product. Potential mutants were identified as having white, or pate blue, roots under these conditions. Out of approximately 79 000 T-DNA mutagenised seedlings screened, two mutants with reduced acid phosphatase staining were further characterised. Both exhibited reduced growth and differences in their P contents when compared to wild-type A. thaliana. The mutant with the most severe phenotype, pho3, accumulated high levels of anthocyanins and starch in a distinctive visual pattern within the leaves. The phenotypes of these mutants are distinct from two previously identified phosphorus mutants (pho1 and pho2) and from an acid phosphatase deficient mutant (pup1) of A. thaliana. This suggested that the screening method was robust and might lead to the identification of further mutants with the potential for increasing our understanding of P nutrition.