Arabidopsis Biological Resource Center

Description:

Line generated in the qrt1-2 background by Agrobacterium-mediated transformation with the pCSA110 T-DNA clone encoding GUS under the control of the postmeiotic, pollen-specific LAT52 promoter, as well as resistance to the herbicide Basta (Glufosinate). Individual BastaR transgenic plants (primary transformants, T1) were self-fertilized to yield T2 seeds. T2 seeds were plated on MS medium containing 50 mg/liter Basta and the percentage of BastaR seedlings was determined. One stage 14 flower from each BastaR plant retained was fixed and stained in X-Gluc overnight at 37°. Pollen tetrads were assayed for meiotic segregation of the LAT52:GUS gene using an inverted microscope. Transmission of the T-DNA through the male gametophyte was tested by pollinating stage 14 ms1 flowers (male-sterile, Col ecotype) with mature pollen from hap+/- plants. Transmission through the female gametophyte was monitored by emasculating stage 12 hap+/- flowers and pollinating them 24-40 hr later with qrt1-2 (Col ecotype) pollen. F1 seed was plated on Basta-containing MS media and seedlings were scored for resistance or sensitivity. Pollen behavior was examined after crossing three plants from each hap line to three or more ms1 pistils and allowing 12 hr for pollen tube growth. Pistils were excised and mounted on double-sided tape; ovary walls were then removed under a dissecting scope. Pistils were prepared and imaged using a Zeiss Axioskop. For closer examination of pollen tube behavior on the ovule, pistils were stained in 0.1% Congo red following incubation in X-Gluc, allowing analysis by confocal laser scanning microscopy. Genomic sequences flanking the right and left T-DNA borders were amplified with thermal asymmetric interlaced (TAIL) PCR.

Growth Requirement: none

Marker:

Background: Col (Columbia); Ler (Landsberg erecta)

ABRC Comment:

Format Shipped: 100 seeds per vial

Base / Commercial Price: $15 / $120