Mapping of genes for root characters in common bean, Marcela Rojas\ Steve Beebe^ Fabio Pedraza^ Fernando Muoz^

Douglas Beck^ Jonathan Lynch^ Sieglinde Snapp^, Joe Tohme^. ^Universiiy of California, Riverside; ^CIAT, Cali, Colombia; ^ 3134 S. Hwy. 99, Merced, CA 95340; "^Penn State University, State College, PA; ^Rockefeller Foundation, Malawi. Introduction Common bean yields are depressed by poor soil fertility in a large part of the tropics and subtropics where much of the world production occurs. Phosphorus (P) and nitrogen are the two elements that are most often deficient but poor availability of potassium, calcium, boron, sulfiu- and other micronutrients can also limit production. Fertilization is only a partial solution to poor soil fertility due to fixation of phosphorus by soil clays and leaching of nitrogen through the soil under high rainfall conditions. In any case, whether nutrients are supphed through native soil fertihty or through fertilizers, the plant requires a healthy and vigorous root system to absorb nutrients. Genetic differences in root systems exist among genotypes and can be exploited to enhance nutrient recovery. The purpose of this study was to identify DNA markers for QTL loci that contribute to greater root length and nutrient absorption, with special reference to P absorption. Materials and methods Two genotypes contrasting for vigor of root system were identified. DOR 364 is a small seeded Mesoamerican cultivar with a course, non-fibrous root system and relatively poor yields imder P deficiency. G1983 3 (Chaucha Chuga) is a large seeded Peruvian landrace of the Andean gene pool; G19833 has an extensive and fibrous root system under both P stress and P sufficient conditions. Recombinant Inbred Lines (RILs) were developed from these two parents and 71 of these were planted with 3 repetitions under P sufficient and P stress conditions in an andisol soil in Darin, Colombia. Root systems were extracted from the soil manually and were weighed. A representative sample of each root system was taken for scarming with the Delta-T-Scan program which permits analyzing total root length, proportion of roots in several diameter classes and average root diameter. Data on the root sample was extrapolated to the whole root system on a weight/weight basis. Standard physiological parameters of root, shoot and total biomass were obtained, as well as total P absorption. Seed size was measured as grams per 100 seed on samples harvested under high P. Probes obtained fi'om the University of Florida and the University of California-Davis were assayed on the two parents in combinations with five different restriction en2ymes. Out of 101 probes, 80% were polymoiphic with one or more enzyme. Seventy probes were assayed on the RILs to create an RFLP framework. Additionally, 44 AFLP and 80 RAPD markers were mapped. A total of 194 markers were analyzed by the Q-Gene program obtained from Cornell University to determine linkage of markers to QTL loci controlling root characters, P absorption and seed size. Results and discussion As many as 22 loci were identified which explained at least 5% of the variability in root length at low P. Of these, the six most important QTL explained 39% percent of the variability in the multiple regression model This figure could not be improved significantly by including more QTL. In any case, if QTL are to be selected in practical breeding programs, it is important to assure that QTL under selection are in fact having the desired impact on nutrient absorption. Therefore the effect of the QTL on P absorption was also studied. Only 5 of the QTL affecting root length at low P also had a sigiificant effect on P absoiption. Curiously enough, the QTL that explained the greatest effect in P absorption at low P (15%) was not related to root length at all. This suggests that genes affecting p^ameters other than root length are also influencing P absorption. These parameters might be root diameter, root hair length, or P mobilization mechanisms related to rhizosphere acidification or exogenous root phosphatase.

The mapping of genes for seed size revealed that QTL that affected root length were often associated with seed size as well. Five such cases were observed. This suggests that some general gene effects associated with larger plant organs are being detected. This may be related directly or indirectly to larger cell size. In Tab. 1 are presented the effects of six loci on four traits: root length, biomass, phosphorus uptake, and seed size. Variabihty explained by the multiple regression model was greater in low P than high P for most traits. Conclusions It has been possible to map QTL in common bean that are associated with greater root length. Several of these are associated with P absorption and therefore may be of interest to incorporate into breeding programs. Some QTL affecting root length appear to be associated with general effects on organ size. However, the one QTL that most affected P absorption was not related to root length and may reflect other mechanisms of P acquisition.

Table 1, Variability (R^) in four traits of common bean explained by molecular markers

M RKER

LINKAGE GROUP

ROOT LENGTH PP+

BIOM ASS PP+

PUPTAKE PP+

100 SEED wt P+

K 066c Bng204 Bng096 Bngl04 H 18Sw Bng091 MODEL

ch rom B ch rom A ch rom D ch rom G ch rom J Unlinked A djusted R Sq

0.16 0.5 0.12 0.11 0.11 0.11 0,39

0.06 0.05 0.01 0.00 0,04 0.16 0.13

0.11 0.14 0.00 0.11 0.08 0.03 0.21

0.05 0.07 0.00 0.04 0.08 0.05 0.08

0.10 0.09 0.00 0.06 0,04 0.01 0.14

0.01 0.01 0.01 0.02 0.08 0.02 0.02

0.05 0.04 0.00 0.11 0.18 0.03 0.16