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Puroindoline B Limits Binding of Puroindoline A to Starch and Grain Softness

Dep. of Plant Sciences and Plant Pathology, Montana State Univ., Bozeman, MT 59717-3150

View larger version (66K): [in a new window]   Fig. 1. Cleaved amplified polymorphism sequence (CAPS) test used to distinguish between Pinb-D1a (Heron allele) and Pinb-D1b (Hi-Line allele) at Ha. BsrBI digests the Pinb-D1a PCR product once (340- and 129-bp products) and the Pinb-D1b PCR product twice (245-, 129-, and 95-bp products). The "undigested" lane was loaded with undigested Pinb-D1a PCR product. MW represents a 1-kb molecular weight standard (Promega, Madison, WI).

View larger version (20K): [in a new window]   Fig. 2. Frequency distribution for grain hardness of progeny derived from crosses of Heron (Pina-D1a/Pinb-D1a) with Hi-Line (Pina-D1a/Pinb-D1b) transgenic isolines with added Pina (HGA), Pinb (HGB), or both Pina and Pinb (HGAB), where progeny were determined homozygous for the presence of the added transgene and the Pin-D1a allele inherited from Heron. From top to bottom, the three histograms compare the following: (A) HGA1 vs. HGA3, which express additional Pina, (B) HGB5 vs. HGB12, which express additional Pinb, and (C) HGAB18 vs. HGAB3, which express additional Pina and Pinb. SKCS values were the mean of F2 plant and replicated 3-m rows grown in rainfed and irrigated environments.

View larger version (33K): [in a new window]   Fig. 3. Extraction of (A) TX114 soluble and (B) friabilin or starch-bound puroindoline proteins from six transgenic lines derived from crosses of Heron (Pina-D1a/Pinb-D1a) with Hi-Line (Pina-D1b/Pinb-D1b) transgenic isolines with added Pina (HGA), Pinb (HGB) or both Pina and Pinb (HGAB) and hard and soft wheat controls. The PINA and PINB proteins are marked with arrows. All HG line extracts were prepared from F4 seeds homozygous for the Pin transgene and the native Pinb-D1a allele inherited from Heron. The Heron ladder serves as a loading control with all other samples loaded at 1x for comparison to Heron.