Inheritance of Anthracnose Resistance in the Common Bean Differential Cultivar G 2333 and Identification of a New Molecular Marker Linked to the Co-42 gene

The inheritance of anthracnose resistance of the common bean ( Phaseolus vulgaris L.) differential cultivar G 2333 to Colletotrichum lindemuthianum races 73 and 89 was studied in crosses with the susceptible cultivar Rudá. The segregation ratios of 15 : 1 in the F₂ and 3 : 1 in the backcrosses to Rudá indicate that for each of the races tested there are two independent resistance loci in G 2333. A random amplified polymorphic DNA (RAPD) molecular marker (OPH18_1200C) linked in resistance to race 73 was identified in a BC₃F_2:3 population derived from crosses between Rudá and G 2333. A RAPD molecular marker OPAS13_950C, previously identified as linked to gene Co-4² , was also amplified in this population. Co-segregation analyses showed that these two markers are located at 5.6 (OPH18_1200C) and 11.2 (OPAS13_950C) cM of the Co-4² gene. These markers were not present in BC₁F_2:3 plants resistant to race 89 indicating that this population carries a different resistance gene. DNA amplification of BC₁F_2:3 plants with RAPD molecular marker OPAB_450C, previously identified as linked to gene Co-5 , indicated that this gene is present in this population.     

Allelism Tests between the Rust Resistance Gene Present in Common Bean Cultivar Ouro Negro and Genes Ur-5 and Ur-11

The pathogenic variability of the fungus Uromyces appendiculatus is an obstacle for the creation of rust‐resistant common bean (Phaseolus vulgaris L.) varieties. Gene pyramiding is an alternative strategy for the development of varieties with durable resistance. However, to reach this goal it is important to identify different genes with ample resistance spectra. Cultivars Ouro Negro, Mexico 309 and Belmidak RR‐3 have been shown to be resistant to several rust races identified in the state of Minas Gerais, Brazil. Ouro Negro is the only rust resistance source being used in the BIOAGRO/Universidade Federal de Viçosa (UFV) breeding programme, which aims at pyramiding resistance genes in the ‘carioca‐type’ cultivar Rudá. It would be also interesting to use Mexico 309 (Ur‐5) and Belmidak RR‐3 (Ur‐11) in the breeding programme. However, there is no available information on the possible allelic relationships between the Ouro Negro resistance gene and Ur‐5 and Ur‐11. This work aimed at: (1) determining the allelic relationship between the Ouro Negro resistance gene and Ur‐5 and Ur‐11; and (2) evaluating a random amplified polymorphic DNA (RAPD) marker previously reported as being linked to Ur‐11, in populations from crosses between Belmidak RR‐3 and Rudá. The allelism tests confirmed that the Ouro Negro rust resistance gene is distinct from Ur‐5 and Ur‐11 and the molecular analyses confirmed that the RAPD marker can be used in our breeding programme to develop ‘carioca‐type’ cultivars with the Ur‐11 gene.     

Multiline as a Strategy to Reduce Damage Caused by Colletotrichum lindemuthianum in Common Bean

The aim of the study was to verify whether a mixture of lines containing equal amounts of seven lines of Carioca‐type common bean, all agronomically uniform but each presenting different patterns of resistance to Colletotrichum lindemuthianum, would be less damaged by anthracnose than the individual pure lines. Plants cultured in experimental plots in Lavras, Minas Gerais, Brazil, in the dry harvest seasons of 2007 and 2008 were inoculated with a mixture of races 65, 81, 87, 89 and 337 of the pathogen, and the severity of anthracnose was evaluated at 10‐ day intervals commencing 12 days after inoculation. The progress of the disease was estimated from the coefficients of the linear regression equations (b₁) and from the areas under the disease progress curves (AUDPC). The mean grain yields were determined in both experimental periods. The value of b₁ for the multiline was not different from that presented by the resistant line MA‐II‐22 and indicated a slower progress of the disease over time compared with susceptible lines. There were no differences in AUDPC values between the multiline and the resistant lines. The multiline presented a grain yield that was similar to those of the most productive lines even though susceptible lines comprised more than 28% of the mixture and such lines showed the lowest yields of grain. It is concluded that the use of the mixture of lines represents a good strategy for reducing the progress of anthracnose in the field and, as a consequence, reducing loss of grain yield.     

Photosynthetic Gas Exchange in Common Bean Submitted to Foliar Sprays of Potassium Silicate,Sodium Molybdate and Fungicide and Infected with Colletotrichum lindemuthianum

This study investigated whether foliar sprays of potassium silicate (KSi), sodium molybdate (NaMo) or a combination of both (KSi + NaMo), with or without the fungicide azoxystrobin (Azox), could reduce anthracnose symptoms, improve photosynthesis and increase yield. Two 2 × 4 factorial experiments, consisting of untreated or fungicide‐treated plants sprayed with KSi, NaMo or KSi + NaMo were arranged in a randomized block design with three replications. The treatments were as follows: (i) KSi; (ii) NaMo; (iii) KSi + NaMo; (iv) Azox; (v) Azox + KSi; (vi) Azox + NaMo; (vii) Azox + KSi + NaMo; and (viii) control (no KSi, NaMo or Azox). The KSi, NaMo and Azox treatments were applied at the rates of 35 g/l, 90 g/ha and 120 g ai/ha, respectively. KSi was applied at 20, 27, 40 and 55 days after sowing (das). NaMo was applied only at 27 das, whereas the fungicide was applied at 27, 40 and 55 das. The plants were inoculated with Colletotrichum lindemuthianum at 23 das. The anthracnose severity was reduced by 64.25% and yield increase by 156.2% in plants sprayed with fungicide compared with non‐sprayed ones. The KSi, NaMo and NaMo + KSi applications reduced anthracnose severity by 31.8, 16.1 and 37.9%, respectively, while the yield increased by 16.8, 18.9 and 63.9%, respectively. There was no difference between treated and non‐treated plants with KSi with respect to the leaf gas exchange parameters C_i, E and g_s. However, A significantly increased by 16.9% in plants treated with Azox. The A was not affected by KSi or NaMo spray; however, it was significantly increased by 12.5% after spraying with NaMo + KSi. In conclusion, bean plants treated with Si and Mo were associated with a decrease in anthracnose as well as an enhancement in photosynthesis activity under field conditions.     

Gibberellic Acid Treatment Reduces the Tolerance of Field-Grown Common Bean to Leaf Removal

I studied the influence of gibberellic acid (GA₃) treatment in a field population of common bean on plant tolerance to leaf removal. Individual bean seedlings were treated with a foliar application of 10 μM GA₃ on day 7 and day 14 after emergence, which led to a significant increase in height in GA₃-treated plants. Twenty-eight days after emergence, either zero, one, two, or three leaflets from each trifoliate leaf were removed from each of 20 GA₃-treated and 20 control plants. All pods were harvested from each plant after plants became senescent 6 weeks later. Multivariate analyses revealed that leaf removal produced significant reductions in several yield components in both GA₃-treated and control plants, although the effects were not pronounced until at least two leaflets from each trifoliate leaf (67% of the total leaf area) were removed. However, GA₃-treated plants suffered greater reductions in total pod wall mass and total seed number than control plants after 33 and 67% leaf area removal. These results indicate that GA₃ treatment may have altered the assimilatory capacity or resource allocation pattern of treated plants in such a way as to decrease their ability to tolerate leaf removal, a negative consequence of the hormonal alteration of traits important to plant compensation for biotic stressors. Received December 6, 1996; accepted March 5, 1997     

Association of Dominant and Recessive Genes Confers Anthracnose Resistance in Stem and Leaves of Common Bean

Different genes might be involved in Colletotrichum lindemuthianum resistance in leaves and stem of common bean. This work aimed to study the genetic mechanisms of the resistance in the leaf and stem in segregating populations from backcrosses involving resistant cultivar AN 910408 and susceptible cultivar Rudá inoculated with spore suspensions of C. lindemuthianum race 83. Our results indicate that two genes which interact epistatically, one dominant and one recessive, are involved in the genetic control of leaf anthracnose resistance. As for stem anthracnose resistance, two genes also epistatic, one dominant and one recessive, explain the resistance to C. lindemuthianum race 83. The recessive gene is the same for leaf and stem resistance; however, the dominant genes are distinct and independent from each other. The three independent resistance genes of AN 910408 observed in this work could be derived from Guanajuato 31.     

Common Bean Reaction to Fusarium oxysporum f. sp. Phaseoli, the Cause of Severe Vascular Wilt in Central Africa

During the September‐December season of 1990, severe symptoms of Fusarium wilt were for the first time observed on a popular climbing bean (Phaseolus vulgaris L.) cultivar. G 2333. introduced within the previous 5 years. Seventy‐three bean genotypes were screened for resistance lo the disease, using artificial inoculation. The effect of inoculation density on the reaction of four selected genotypes was also investigated. Of the 29 climbing bean genotypes evaluated, 19 were resistant, including 11 of the 15 pre‐release or released cultivars. Of the 44 bush bean cultivars evaluated, 28 were resistant, five were intermediate and 11 were susceptible. All susceptible cultivars showed vascular discoloration. In both susceptible and resistant genotypes, the fungus spread almost equally from the entry points in inoculated roots to the base of the plants, but colonization and vertical spread within the vascular system were markedly less in resistant than in susceptible cultivars. At 20 and 30 cm above soil level, the fungus was only recovered from susceptible cultivars. Increasing inoculum density from 10² to 10⁷ conidia/ml did not affect the resistance of cultivars RWR 950 and G 685 but. in the susceptible cultivars G 2333 and MLB‐48‐89 A. it resulted in early appearance, high incidence and severity of the disease.     

Bean (Phaseolus vulgaris L.) protoplasts as a model system to study the expression and stability of recombinant seed proteins

A suitable protocol for the transient expression of seed protein genes in protoplasts derived from cell suspension cultures of common bean has been established. Preliminary analyses of cultures to verify the synthesis of phaseolin – actively accumulated by the starting tissue, the developing cotyledon – showed that the protein was no longer synthesised after 5 days of culture. Transient expression of a phaseolin sequence, driven by a constitutive promoter, resulted in the accumulation of the correctly glycosylated and assembled protein. This system, when compared to tobacco protoplasts, largely avoids phaseolin fragmentation and the presence of contaminant polypeptides in the immunoprecipitates. Therefore, bean protoplasts are a good system to study the expression of wild-type as well as in-vitro-modified bean seed proteins. Received: 26 November 1996 / Revision received: 10 February 1997 / Accepted: 15 February 1997     

Effect of Cytokinin 4-PU-30 on the Lipid Composition of Water Stressed Bean Plants

Fourteen days-old bean plants, grown on sand with Knop's nutrient solution were subjected to water stress (three days without irrigation). The stress led to a decrease in almost all lipid classes except phospholipids in the primary leaves. The content of palmitic acid increased, and that of the linolenic acid decreased. An increase of hexadecenoic acid in phospholipids was also observed. Rewatering for 24 h led to the recovery of the stressed plants including that of the photosynthetic apparatus, but the changes in the lipid composition were insignificant. The spraying of the plants before and after the water stress with 5 × 10-6 M solution of the phenylurea cytokinin 4-PU-30 alleviated negative effect of water stress on the lipid membrane composition permitting the plants to resist the harmful environment. This revised version was published online in July 2006 with corrections to the Cover Date.     

On the Relationship between the Activity of O-Methyltransferase and the Content of Lignin in Various Organs of Kidney Bean

The activity of O-methyltransferase (OMT) and the content of lignin in various organs of 12- to 14-day-old seedlings of kidney bean (Phaseolus vulgarisL., cv. Saksa) were determined. The epicotyl and hypocotyl were characterized by the highest OMT activity, and the leaves, by the lowest OMT activity. At the same time, the lignin content gradually increased in the following order: leaves < epicotyl < hypocotyl < roots. Thus, in kidney bean seedlings, there is no direct relationship between the OMT activity and the lignin content. It is suggested that lignin biosynthesis is limited by biosynthetic steps other than O-methylation of caffeic and 5-hydroxyferulic acids affected by OMT.     

Sources of Resistance to Anthracnose in Traditional Common Bean Cultivars from Paraná, Brazil

Pathogenicity of physiologically distinct races of Colletotrichum lindemuthianum originating from Andean (races 7, 19 and 55) and Mesoamerican (races 9, 31, 65, 69, 73, 81, 89, 95 and 453) locations of the new world were evaluated on 26 landrace genotypes of common bean (Phaseolus vulgaris L.) from Paraná State, Brazil. Races 7 (Andean), 65, 73 and 89 (Mesoamerican) were the most pathogenic, while race 31 (Mesoamerican) was the least pathogenic. Most of the landrace genotypes evaluated (88%) were resistant to race 31, except Carioca 3, Preto 1 and Preto 2. In addition, about 50% of the landrace genotypes had resistance to races 9, 19, 55 and 453; and about 30% to races 7, 65, 69, 73, 81, 89 and 95. The resistance index, which measured the pathogenicity response averaged across all the physiologically distinct Andean and Mesoamerican races of C. lindemuthianum, of the landrace genotypes ranged from 8% to 83%. The most resistant cultivars were Carioca Pintado 1, Carioca Pintado 2, Jalo Vermelho and Jalo de Listras Pretas. In contrast, the most susceptible cultivars were Jalo Pardo, Jalo Pintado 1 and Bolinha that showed resistance only to the least pathogenic race 31. These results indicated that many of the common bean landrace cultivars evaluated have genes that could be useful in breeding programmes to enhance resistance to Andean and Mesoamerican races of C. lindemuthianum.     

Resistance of Common Bean (Phaseolus vulgaris) to Bacterial Wilt Caused by Curtobacterium flaccumfaciens pv. flaccumfaciens

Bacterial wilt caused by Curtobacterium flaccumfaciens pv. flaccumfaciens is an important new disease of common bean (Phaseolus vulgaris) in western Canada. Both yellow and orange variants of the pathogen were found in the region. A controlled environment study was conducted to assess 124 common bean cultivars and lines from eight market classes for resistance to the yellow and orange variants of the pathogen, using the hilum injury/seed inoculation method. Results of the screening tests showed significant (P < 0.05) differences in resistance to bacterial wilt among the cultivars or lines. The great northern line L02E317, the great northern cultivar Resolute and pinto lines L02B662 and 999S‐2A, were highly resistant to both variants of the pathogen, with disease severity indices of 0 on a rating scale of 0 (no wilt symptoms) to 5 (dead seedling). Resistant cultivars or lines were found among black, great northern, pink, pinto, small red and Flor de Mayo bean market classes. The study concludes that new bacterial wilt‐resistant germplasm exists among Canadian bean cultivars and lines, and constitutes a valuable resource for breeding common beans for resistance to both yellow and orange variants of C. flaccumfaciens pv. flaccumfaciens.     

菜豆泛肽基因在生物和非生物胁迫下的表达

差别筛选HgCl2胁迫处理的菜豆（Phaseolus vulgaris L.）幼苗叶片cDNA库,分离出1个重金属胁迫响应基因PvSR52克隆,其cDNA长度为281bp。cDNA和氨基酸序列同源性分析表明PvSR52编码一种多聚泛肽。Southern blot结果表明菜豆泛肽可能由少数基因编码。Northern blot分析表明多聚泛肽叶片中表达较少;重金属Hg、Cd和As等、过量的Zn和Cu及高温、病毒侵染和水杨酸等环境胁迫均能强烈地刺激其在叶片中的表达。推测泛肽水解系统在提高植物的抗塑性方面有重要作用。     

Effect of Salt Stress on Carbon Metabolism and Bacteroid Respiration in Root Nodules of Common Bean (Phaseolus vulgaris L.)

Abstract: In the present work, we examined the effect of salinity on growth, N fixation and carbon metabolism in the nodule cytosol and bacteroids of Phaseolus vulgaris, and measured the O₂ consumption by bacteroids incubated with or without the addition of exogenous respiratory substrates. The aim was to ascertain whether the compounds that accumulate under salt stress can increase bacteroid respiration and whether this capacity changes in response to salinity in root nodules of Phaseolus vulgaris. The plants were grown in a controlled environment chamber, and 50, 100 mM or no NaCl (control) was added to the nutrient solution. Two harvests were made, at the vegetative growth period and at the beginning of the reproductive period. The enzyme activities in the nodule cytosol were reduced by the salt treatments, while in the bacteroid cytosol the enzyme activities increased at high salt concentrations at the first harvest and for ADH in all treatments. The data presented here confirm that succinate and malate are the preferred substrates for bacteroid respiration in common bean, but these bacteroids may also utilize glucose, either in control or under saline conditions. The addition of proline or lactate to the incubation medium significantly raised oxygen consumption in the bacteroids isolated from plants treated with salt.     

Phototropic Response of the Bean Pulvinus: Movement of Water and Ions

Segmental analysis of the laminar pulvinus of Phaseolus vulgaris L. showed that its phototropic curvature is accompanied by efflux of inorganic ions and water from its contracting sector and a comparable influx into its expanding one. All the major ions, except Na⁺, contributed to this transport, suggesting that the response to light involves changes in the driving force, or conductivity of a wide range of solutes. During the curvature, K⁺ and CI^? made the greatest and equivalent contributions to efflux, but only Cl^? exhibited a matching influx into the expanding sector, while K⁺ influx was much less. Use of the cell pressure probe showed that, as the laminar angle of elevation changed between ?40° to +40°, turgor pressure in the expanding motor cells increased by 0.48 MPa and decreased in the contracting cells by 0.32 MPa. Picoliter osmometry of single-cell samples showed that during this movement vacuolar osmotic pressure remained constant. Thus, changes in turgor pressure resulted from changes in apoplastic, rather than the protoplastic osmotic pressure. Volumetric modulus of elasticity of pulvinar motor cells is very low, showing that their walls are very elastic. These properties increase the effectiveness of converting osmotic work into the large-scale, reversible volume changes responsible for leaf movements.     

Biological Control of Bacterial Wilt of Bean Using a Bacterial Endophyte, Pantoea agglomerans

Indoor studies were conducted to determine the potential use of Pantoea agglomerans isolate LRC 8311 as a biocontrol agent for control of bacterial wilt of bean caused by Curtobacterium flaccumfaciens pv. flaccumfaciens. Soaking seeds of great northern bean cv. US1140 in a suspension of 3 × 10⁸ cfu/ml P. agglomerans resulted in thorough endophytic colonization of the entire bean seedling from root to apical stem after 7 days, regardless of whether the inoculated seeds were hilum injured or not. Colonization of seedlings by P. agglomerans increased seedling height after 10 days, and had no negative effect on seedling emergence. Treatment of hilum‐injured bean seeds of great northern bean cv. US1140 or navy bean cv. Morden003 with a mixture of P. agglomerans + C. flaccumfaciens pv. flaccumfaciens resulted in a high rate of colonization of seedlings by P. agglomerans, reduced frequency of infection by C. flaccumfaciens pv. flaccumfaciens, improved seedling emergence and height, and reduced disease severity, compared with seeds treated with the wilt pathogen alone. Application of P. agglomerans as a soil drench 24 h after planting was also effective in suppressing bacterial wilt in some instances, but was generally not as effective as seed treatment. The study suggests that seed treatment with P. agglomerans may be an effective and practical method for control of bacterial wilt of bean.     

菜豆重金属胁迫响应基因:cDNA克隆及其表达分析

通过差别筛选HgCl2胁迫下的菜豆叶片cDNA库,分离出7组不同的cDNA克隆（Phaseolusvulgarisstress-relatedprotein,PvSR1～7）。cDNA序列和同源性分析结果表明：PvSR1编码富含脯氨酸细胞壁蛋白（PRP）,PvSR2和PvSR7编码新的HgCl2胁迫相关蛋白,PvSR3编码脱水蛋白（dehydrin）,PvSR4编码病原相关（PR）蛋白,PvSB5编码polyubiqui－tin,PvSR6编码DuaJ－like蛋白。HgCl2胁迫可强烈地请导PvSR2和PR蛋白基因的表达,并能提高PRP,、dehydrinlike和polyubiq－uitin基因的转录水平。这些蛋白质共同作用可能对维持细胞的正常代谢和抵抗重金属胁迫方面有重要作用。