Abstract
Phosphatidylinositol 3-kinase has been reported to be important for normal plant growth. To characterize the role of the enzyme further, we attempted to isolate Arabidopsis (Arabidopsis thaliana) plants that do not express the gene, but we could not recover homozygous mutant plants. The progeny of VPS34/vps34 heterozygous plants, harboring a T-DNA insertion, showed a segregation ratio of 1:1:0 for wild-type, heterozygous, and homozygous mutant plants, indicating a gametophytic defect. Genetic transmission analysis showed that the abnormal segregation ratio was due to failure to transmit the mutant allele through the male gametophyte. Microscopic observation revealed that 2-fold higher proportions of pollen grains in heterozygous plants than wild-type plants were dead or showed reduced numbers of nuclei. Many mature pollen grains from the heterozygous plants contained large vacuoles even until the mature pollen stage, whereas pollen from wild-type plants contained many small vacuoles beginning from the vacuolated pollen stage, which indicated that vacuoles in many of the heterozygous mutant pollen did not undergo normal fission after the first mitotic division. Taken together, our results suggest that phosphatidylinositol 3-kinase is essential for vacuole reorganization and nuclear division during pollen development.
Highlights
Phosphatidylinositol 3-kinase has been reported to be important for normal plant growth
It is essential for normal growth, as demonstrated by the expression of AtVPS34 antisense constructs, which leads to second-generation transformed plants with very severe defects in growth and development (Welters et al, 1994)
We discovered that the self-fertilized heterozygous plants produced progeny that segregated 1:1 for wild-type and heterozygous plants, and no homozygous mutant plants were recovered
Summary
Phosphatidylinositol 3-kinase has been reported to be important for normal plant growth. To determine which gametophyte caused the abnormal segregation ratio, we tested male and female transmission efficiency (TE) by performing reciprocal crosses between heterozygous and wild-type plants and analyzed the F1 progeny by PCR-based genotyping.
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