Mechanism of Russian wheat aphid resistance.

A.J. van der Westhuizen, X-M. Qian, and A-M. Botha.

The objective of this study is to gain information on the mechanism of resistance that may contribute to the identification of resistance markers and resistance genes.

Resistance is not found to be caused by a constitutive factor, but involves a hypersensitive-type reaction elicited by feeding aphids. The resistance mechanism is induced at a higher level in resistant than susceptible wheat. Indications of the hypersensitive reaction were increased respiration rate; accumulation of phenolic compounds, which may include phytoalexins; and development of necrotic spots. Specific intercellular proteins accumulated in resistant wheat only in reaction to aphid infestation. Some of them were related immunologically to pathogenesis­related proteins. Selective higher increases in chitinase, ß-1, 3-glucanase, and peroxidase activities, were found in resistant wheat within 48 hours after aphid infestation. These increased activities were due to increased de novo synthesis of these enzymes and may serve as quantitative measures of resistance. The injurious effect of Russian wheat aphid infestation on the chloroplast protein Rubisco may be a major contributor to the poor survival abilities of susceptible cultivars.

Although the accumulation of chitinases is selective, ß-1,3-glucanase and peroxidases or their increased activities may serve as biochemical markers of resistance. However, their use would not be substantially time­saving, because plants must be in an early 3-leaf stage and the physical symptoms usually used for selection follow soon afterwards. Regarding the identification of biochemical markers of resistance, our research now concentrates on germlings (24ñ48 h old). Except in the case of unrelated markers to defense, the resistance mechanism has to be elicited first before accumulation of specific proteins or increased enzyme activities could serve as markers. Several potential chemical elicitors are being tested for selective induction of defense related enzyme activities. The intercellular washing fluids of infested resistant wheat were found to contain a selective elicitor. Studies are continuing to determine its nature. The eliciting effects of different chitin isomers have been tested on plantlets and found to be nonspecific. Future studies will include key enzymes of the phenolic synthesis pathway.

These research programs are financially supported by the Wheat Board in agreement with the Small Grain Institute, Bethlehem.


UNIVERSITY OF THE ORANGE FREE STATEóDEPARTMENT OF PLANT PATHOLOGY

Bloemfontein, 9300, South Africa.

Z.A. Pretorius, F.J. Kloppers, and C.M. Bender.

Yield loss study.

The destructive potential of leaf and stem rust on certain bread wheat cultivars currently grown in high-risk areas were confirmed in an artificially inoculated field trial at Greytown during 1995. In comparison with fungicide-treated control plots, mean plot yield was reduced by 45 % (first planting date) and 24 % (second planting). The corresponding reductions in 1,000-kernel weight were 33 % and 18 %, respectively. Leaf and stem rust reached 100S and 80S severity levels on susceptible cultivars.

Improvement of leaf rust resistance.

In cooperation with the Small Grain Institute of the ARC, 1,377 lines derived from the crosses `Karee/Lr29', `Karee/Lr34', `Palmiet/Lr29', and `Palmiet/Lr34' were evaluated for leaf rust reaction in the field. Seed of these lines was harvested for preliminary yield testing in 1996. Lines homozygous for leaf rust resistance currently are being selected from `Karee*6/Lr35' and `Karee*6/Lr37' populations and will be tested accordingly by the Small Grain Institute. Palmiet or Karee with Lr21, Lr32, Lr35, Lr36, Lr41, or Lr42 have been backcrossed four times to the recurrent parent.

Characterization of resistance to wheat leaf rust.

Cornel Bender completed a Master's thesis on Lr12 and Lr13 resistance during 1995. Four Thatcher (Tc) F3 lines (Lr13/Lr12-3, Lr13/Lr12-9, Lr13/Lr12-19, and Lr13/Lr12-40), homozygous for both Lr13 and Lr12, were selected, and their resistance was compared with that of the parents (CT263 [= TC Lr13] and RL6011 [= TC Lr12]), the single gene lines `TC/13-22' and `TC/Lr12-16', and Thatcher. Quantification of aborted penetration showed that inhibition of fungal growth in wheat lines containing Lr12 and/or Lr13 was activated to a certain degree before haustoria were formed. Colony size showed that fungal colonies were generally smaller in lines containing both Lr12 and Lr13 than in the parents, but not necessarily smaller than those in the monogenic line `Tc/Lr13-22'. Host cell necrosis was associated more frequently with infection sites, specifically of pathotype UVPrt2, in the combination lines than in the parents. Hypersensitivity index values indicated that host cell necrosis was more severe following infection of the combination lines with UVPrt2. Quantification of cell wall appositions showed that fewer papillae occurred in Thatcher than in the other host genotypes. The number of haustoria observed per colony did not indicate any clear, repeatable differences between lines. Flag leaf infection types showed that Lr12 is effective to most pathotypes of P. recondita f. sp. tritici occurring in South Africa. Conversely, Lr13 is ineffective to the dominant pathotypes. Ratings on primary and flag leaves, and the resistance components of latent period, uredium density, and uredium size, did not distinguish between the digenic lines and the most resistant parent. In the absence of a pathotype virulent to both genes, the combination lines were highly resistant in the field.

Personnel.

Following a research position, Dr. F.J. Kloppers was appointed as senior lecturer in the department as of 1 January, 1996.

Publications.

Crous PW, Petrini O, Marais GF, Pretorius ZA, and Rehder F. 1995. Occurrence of fungal endophytes in cultivars of Triticum aestivum L. in South Africa. Mycoscience 36:105-111.

Kloppers FJ and Pretorius ZA. 1995. Histology of infection and development of Puccinia recondita f. sp. tritici in a wheat line with Lr37. J Phytopath 143:261-267.

Kloppers FJ, Pretorius ZA, and Van Lill D. 1995 . The influence of Lr29, Lr35 and Lr37 on leaf rust severity, yield loss and quality characteristics in wheat. SA J Plant and Soil 12:55-58.

Pretorius ZA Van Niekerk BD, Kloppers FJ, and Vorster AL. 1995. Managing certain recently named Lr genes in breeding wheat for resistance to Puccinia recondita f. sp. tritici in South Africa. SA J Plant and Soil 12:32-37.


UNIVERSITY OF STELLENBOSCH

Department of Genetics, Stellenbosch 7600, South Africa.

G.F. Marais, R. Prins, A. Antonov, F.L. Middleton, H.S. Roux, and A.S. Marais.

Breeding programs.

Selection programs aimed at developing improved durum cultivars for the lower Orange River irrigation areas and triticale cultivars for the southwestern Cape were continued. No new releases were made, but promising advanced lines with good quality were selected.

Genetic studies.

An attempt to transfer leaf rust resistance genes from Aegilops and Triticum species to common wheat was continued. Resistance genes from 70 accessions were expressed fully in hybrids, and these are now in various stages of backcrossing.

Twenty-nine lines, with deletions in the Lr19 (Indis) translocated chromosome segment, were used to physically map a number of Thinopyrum loci. The relative positions of the marker loci on the translocated segment were determined as: Sd1, Xpsr165, Xpsr105, Xpsr129, Lr19, Wsp-D1, Sr25, Y1, and Y2. When compared to the map of the wheat group 7 chromosomes, the data confirmed the reported homoeology between the Lr19 segment and chromosome arm 7DL of wheat. Also, it is evident that the Lr19 translocation in Indis is very similar to the Lr19 segment in the T4 source and that the former may not derive from Thinopyrum distichum as was previously thought. The mutation data suggest that Sd1 and Lr19 may not be single loci. Polymorphisms produced by eight recombined forms (ph-induced) of the Lr19 (Indis) translocations then were related to the physical map of the region. At least five of the recombinants proved to be double crossover products. Each has lost Sd1, yet they are still associated with mild segregation distortion and sometimes strong self-elimination. However, it is not clear whether the segregation distortion results from additional Sd loci or the altered chromosome structure following relocation of the segments. Evidence was found that Lr19 resistance may be determined by two loci. The successful transfer of a gene for RWA resistance from `Imperial' rye to the 1BL·1RS translocation prompted an attempt to replace the secalin locus on this translocation with wheat chromatin. We attempted the transfer in two ways: (i) by deleting the homoeologous pairing inhibitor, Ph1b, locus (through the use of the ph1b deletion mutant), and (ii) by creating plants that are nullisomic for chromosome 3D (Ph2b absent), but trisomic or tetrasomic 3B (have extra doses of the 3B pairing promotors).

The RWA-resistance gene, Dn5, was found to be located on chromosome arm 7DL (telosomic analysis). The gene segregates independently from the centromere and appears to be located at about 32 map units from the endopeptidase locus, Ep-D1. A dominant gene for stem rust resistance (germplasm line 87M66-2-1), which is derived from T. tauschii accession RL5289, was located on chromosome arm 1DS. The locus appears to be separate from, but linked to, Sr33.

A germplasm line, KS87VP9, which has a dominant gene for male sterility, was obtained from the USDA, and the gene was transferred through backcrossing to a well-adapted spring wheat background. This then was used as one of the parents in a multicross involving 16 wheat parents. The sterility gene was found to work very well, giving regular and complete male sterility to 50 % of the progeny. A procedure was worked out whereby large numbers of selections can be intercrossed with comparative ease, making use of the sterility gene. The multicross F1 will be planted in the field in the coming season and will serve as the starting material for a recurrent selection procedure aimed at improvements in yield, disease resistance, and quality. The aim is to eventually incorporate and evaluate the procedure as part of a pedigree- selection system.

Publication.

Du Toit F, Wessels WG, and Marais GF. 1995. The chromosome arm location of the Russian wheat aphid resistance gene, Dn5. Cereal Res Commun 23(1-2):15-17.