Digested bands detected with non-denaturing polyacrylamide gels stained with either silver (a) or ethidium bromide (b) and agarose gels stained with ethidium bromide (c).

<p>Putative mutations in the pools (1, 2, 3, 4) are identified by the presence of two bands (indicated by white arrows), with sizes adding up to the full length PCR product. (a). Non-denaturing polyacrylamide gel stained with silver; (b). Non-denaturing polyacrylamide gel stained with ethidium bromide; (c). Agarose gels stained with ethidium bromide.</p

Primer sequences of the 6 primers used and the mutation frequency detected.

<p>Primer sequences of the 6 primers used and the mutation frequency detected.</p

Frequency of typical mutations observed among the 2,610 M₂ individuals screened.

<p>Frequency of typical mutations observed among the 2,610 M₂ individuals screened.</p

Type and distribution of induced mutations discovered in <i>Ppd-D1</i> amplicon.

<p>Type and distribution of induced mutations discovered in <i>Ppd-D1</i> amplicon.</p

The mutation density and germination rate in published TILLING populations of different species under different EMS dosage treatments.

*<p>in M₃ plant.</p

Mutation detection and estimation of mutation frequency in three candidate genes of <i>Ppd-D1</i>, <i>Rubisco activase A</i> and <i>Rubisco activase B</i> by TILLING analysis.

a<p>, mutation detected by non-denaturing polyacrylamide gels stained with silver;</p>b<p>, mutation detected by non-denaturing polyacrylamide gels stained with ethidium bromide;</p>c<p>, mutation detected by agarose gels stained with ethidium bromide.</p>*<p>For calculation of the mutation frequency, 100 bp sequences from each end were removed due to the base ambiguity.</p

Mutant phenotypes observed in the M₂ and M₃ wheat plants.

<p>Mutant phenotypes: (a), (i), (j) dwarf and semi-dwarf; (b) single tiller; (c) coleoptile color; (d) seed size; (e), (m) albinism; (f) spike morphology; (g)(h) erect leaf; (k) narrow leaf; (l) strange leaf morphology; (n) disease sensitive; (o) large spikes with short awns; (p) yellow spots on leaves.</p

An example of RAPD banding pattern obtained by primer R1 and primer IT31.

<p>W: wild type; m: mutagenised lines; M: molecular weight standards DL2000.</p

Analysis of mutations identified in the <i>Ppd-D1</i> gene.

*<p>Het, heterozygote; Hom, homozygote.</p><p>PSSM or SIFT scores of mutation lines with star (*) are predicted to be damaging to protein function. Mutation with PSSM score larger than 10 indicates that the mutation is more likely to have a damaging effect on the protein function. Mutation with SIFT score less than 0.05 is predicted to be deleterious.</p