Assessment of the uniformity and stability of grapevine cultivars using a set of microsatellite markers

Solidity of microsatellite markers is a key issue for varietal identification, especially when they are used for legal purposes, what includes their probable future use in the distinctness, uniformity and stability testing of new varieties needed for the granting of Plant Breeders’ Rights. Nine grapevine microsatellites (VVS2, VVMD5, VVMD27, VVMD28, ssrVrZAG29, ssrVrZAG62, ssrVrZAG67, ssrVrZAG83 and ssrVrZAG112), which had previously demonstrated its capacity to discriminate any grapevine variety, have been assessed to evaluate its uniformity and stability. 19 varieties were selected, representative of a high diversity for morphological, agronomical, cultural and historical aspects, as well as for microsatellite allele variability. Then, for each variety, uniformity and stability were evaluated through the analysis of 50 plants from each of three different plots, and five plants from each of seven additional plots. Material from 4,137 plants of 229 plots of the 19 varieties was sampled in seven countries. Of 3,654 plants analyzed with the set of nine microsatellites, 3,299 were of the right variety and used for the survey. An average of 172 individual values was studied for each allele of each microsatellite of each variety, and none differences were detected that could not be explained as technical variations, with the exception of several putative chimeras in two varieties. Of the total of 171 variety x microsatellite combinations, only in one combination (‘Merlot’ x VVMD27) the number of off-types exceeded the threshold allowed. The remaining 170 combinations have been found uniform and stable according to internationally accepted rules.     

Analysis of rice blast resistance gene <Emphasis Type="Italic">Pi</Emphasis>-<Emphasis Type="Italic">z</Emphasis> in rice germplasm using pathogenicity assays and DNA markers

The Pi-z gene in rice confers resistance to a wide range of races of the rice blast fungus, Magnaporthe oryzae. The objective of this study was to characterize Pi-z in 111 rice germplasm accessions using DNA markers and pathogenicity assays. The existence of Pi-z in rice germplasm was detected by using four simple sequence repeat (SSR) markers (RM527, AP4791, AP5659-1, AP5659-5) closely linked to Pi-z, and was verified using pathogenicity assays with an avirulent strain (IE1k) and two virulent races (IB33 and IB49). Among 111 germplasm accessions evaluated, 73 were found to contain the Pi-z gene using both SSR markers and pathogenicity assays. The remaining 38 germplasm accessions were found to be inconsistent in their responses to the blast races IB33, IEIk and IB49 with expected SSR marker alleles, suggesting the presence of unexpected SSR alleles and additional R gene(s). These characterized germplasm can be used for genetic studies and marker-assisted breeding for improving blast resistance in rice.     

Genetic and cytological analysis of a new spontaneous male sterility in radish (<Emphasis Type="Italic">Raphanus sativus</Emphasis> L.)

A male sterile plant appeared in the radish breeding program at the Hubei Academy of Agricultural Sciences, Hubei, China. In its progeny, a two-type (half of plants male sterile, the other half male fertile) line 01GAB was established. An F₂ population of 260 plants from a cross of male-sterile 01GAB and a male fertile line 9802H segregated for male fertility in a 3:1 ratio indicating that fertility was restored by a single dominant gene, here designated RsMs. A PCR-based DNA marker specific to the male fertility Rfob gene in 9802H was absent in 01GAB. Linkage analysis placed the RsMs locus 10.7 cM away from the Rfo locus. In an F₂ population of hybrids between 01GAB and male fertile 9802B, a co-dominant DNA marker for the RSultr3.2A (a radish sulfate transporter gene) locus was linked to the RsMs locus at 1.5 cM suggesting that fertility restoration in 01GAB was located in the region with known male sterility restorers in radish. However, no maintainer for the 01GAB source of male sterility has been identified so far. Cytological observations have shown that the abnormalities in male sterile anthers first appeared in tapetum at the tetrad stage, followed by a hypertrophy of the tapetal cells at the vacuolate microspore period. These results suggest that male sterility in 01GAB is likely to be genetic in nature, or it may represent a new type of the cytoplasmic male sterility.     

Combining different resistance components enhances resistance to <Emphasis Type="Italic">Fusarium</Emphasis> head blight in spring wheat

Fusarium head blight (FHB) is a devastating disease in wheat throughout the world. FHB resistance consists of two components: resistance to initial infection (type I) and resistance to spread within infected spikes (type II). Current wheat breeding programs for FHB focus on type II resistance, which limits pathogen spread but may not be sufficiently durable. To combine type I with existing type II resistance, 113 F₉-derived recombinant inbred lines (RILs) were developed from a cross between three wheat genotypes Frontana, W9207, and Alsen. The RILs were evaluated for resistance to initial infection, FHB spread within spike, kernel damage, and deoxynivalenol (DON) content in two independent greenhouse experiments in 2006 and 2007. Among the 113 RILs, 20% lines showed ≤10% initial disease severity (IDS) and ≤11 to 30% final disease severity (FDS), and 19% had DON content ≤5 μg/g. Approximately 11% of the RILs showed tendency of higher resistance (as exhibited by lower IDS, FDS, and DON content) than the resistant parents. The 42 of the FHB-resistant RILs were analyzed with seven simple sequence repeat (SSR) markers or microsatellites known to be linked to FHB resistance. Approximately half of the RILs had molecular markers linked to both types of FHB resistance indicated the presence of type I and II resistance alleles in the RILs. The resistant RILs identified in this study should be useful for the future improvement of FHB resistance in spring wheat.     

Successful induction of trigenomic hexaploid <Emphasis Type="Italic">Brassica</Emphasis> from a triploid hybrid of <Emphasis Type="Italic">B.</Emphasis><Emphasis Type="Italic">napus</Emphasis> L. and <Emphasis Type="Italic">B. nigra</Emphasis> (L.) Koch

A triploid hybrid with an ABC genome constitution, produced from an interspecific cross between Brassica napus (AACC genome) and B. nigra (BB genome), was used as source material for chromosome doubling. Two approaches were undertaken for the production of hexaploids: firstly, by self-pollination and open-pollination of the triploid hybrid; and secondly, by application of colchicine to axillary meristems of triploid plants. Sixteen seeds were harvested from triploid plants and two seedlings were confirmed to be hexaploids with 54 chromosomes. Pollen viability increased from 13% in triploids to a maximum of 49% in hexaploids. Petal length increased from 1.3 cm (triploid) to 1.9 cm and 1.8 cm in the two hexaploids and longest stamen length increased from 0.9 cm (triploid) to 1.1 cm in the hexaploids. Pollen grains were longer in hexaploids (43.7 and 46.3 μm) compared to the triploid (25.4 μm). A few aneuploid offsprings were also observed, with chromosome number ranging from 34 to 48. This study shows that trigenomic hexaploids can be produced in Brassica through interspecific hybridisation of B. napus and B. nigra followed by colchicine treatment.     

Three AFLP markers tightly linked to the genic male sterility <Emphasis Type="Italic">ms</Emphasis><Subscript><Emphasis Type="Italic">3</Emphasis></Subscript> gene in chili pepper (<Emphasis Type="Italic">Capsicum annuum</Emphasis> L.) and conversion to a CAPS marker

Genic male sterility (GMS) has long been used as a tool for hybrid seed production in chili pepper (Capsicum annuum L.). We developed DNA markers linked to the GMS ms ₃ gene in a segregating population using bulked segregant analysis (BSA) and amplified fragment length polymorphism (AFLP) techniques. The segregating population was subjected to BSA-AFLP with 512 primer combinations. Three AFLP markers (Eagg/Mccc₂₇₆, Eagc/Mctt₁₇₈, and Ecag/Mtgc₂₀₄) were identified as tightly linked to the ms ₃ locus. Among them, we converted the AFLP marker Ecag/Mtgc₂₀₄ to the cleavage amplified polymorphic sequence (CAPS) marker, named GMS3-CAPS, based on sequencing analysis of internal and flanking regions for the markers between male-fertile and sterile plants. This marker will be useful for pepper breeding using the GMS system.     

Quantitative trait loci influencing fruit-related characteristics of tomato grown in high-temperature conditions

The objective of this study was to identify molecular markers linked to fruit-related traits in the tomato subjected to high temperatures. In total, 160 F₂ plants derived from a cross between a heat-tolerant breeding line, CL5915-93D4-1-0-3 (Solanum esculentum), and a heat-sensitive wild accession, L4422 (S. pimpinellifolium), were grown in a greenhouse. Six traits including fruit number, fruit weight, brix, seed number, fruit setting, and flower number were scored. The distributions of fruit number, fruit set, flower number, and seed number were skewed towards heat susceptibility which is known to be characteristic of L4422. Polymorphic bands were generated by PCR-derived methods of RAPD, ISSR and AFLP Polymorphism, the segregation ratio, and distribution over the genome of the above 3 markers were compared. Ten linkage groups, ranging 20.6–151.6 cM in size, were constructed with 62 informative markers spanning a total of 776.3 cM. Fruit-related quantitative trait loci (QTLs) were non-randomly distributed in the tomato genome. For the 6 traits investigated, 21 QTLs were dispersed on linkage groups 2–5. The genetic effects of the various QTLs were differently exhibited, in our study we have respectively found from 10.5% to 30.2% of the variation explained by the QTL for flower number (FRN4) and brix (BX2). Thirteen QTL-mapped markers were unique to 1 trait, and 4 markers were linked to more than 1 trait. Among them, QTLs linked to the I868-470 marker had effects on fruit weight and brix, and a significant positive correlation between these 2 traits was noted (r = 0.35, P < 0.05). Thus, the I868-470 marker may have the potential for simultaneous selection of high fruit weight and brix. These markers also allowed us to align genome linkage maps across distantly related species and to reveal the co-localization between these QTLs and major genes.     

Comparative study of conventional and artificial neural network-based ETo estimation models

Accurate estimation of reference crop evapotranspiration (ETo) is required for several hydrological studies and thus, in the past, a number of ETo estimation methods have been developed with different degree of complexity and data requirement. The present study was carried out to develop artificial neural network (ANN) based reference crop evapotranspiration models corresponding to the ASCE’s best ranking conventional ETo estimation methods (Jensen et al. ASCE Manual and Rep. on Engrg. Pract. no. 70, 1990). Among the radiation methods, FAO-24 radiation (or Rad) method for arid and Turc method for humid region, and among the temperature methods, FAO-24 Blaney–Criddle (or BC) method were studied. The ANN architectures corresponding to the above three less data-intensive methods were developed for four CIMIS (California Irrigation Management Information System) stations, namely, Davis, Castroville, Mulberry, and West Side Field station. The comprehensive ANN architecture developed by Kumar et al. (J Irrig Drain Eng 128(4):224–233, 2002) corresponding to Penman–Monteith (PM) ETo for Davis was also tried for the other three stations. Daily meteorological data for a period of more than 10 years (01 January 1990 to 30 June 2000) were collected from these stations and were used to train, test, and validate the ANN models. Two learning schemes, namely, standard back-propagation with learning rate of 0.2 and standard back-propagation with momentum having learning rate of 0.2 and momentum term of 0.95 were considered. ETo estimation performance of the ANN models was compared with the FAO-56 PM method. It was found that the ANN models gave better closeness to FAO-56 PM ETo than the best ranking method in each category (radiation and temperature). Thus these models can be used for ETo estimation in agreement with climatic data availability, when not all required climatic variables are observed.     

Optimal utilisation of natural resources for agricultural sustainability in rainfed hill plateaus of Orissa

The climatic data for 17 years from 1988 to 2004 of the rainfed hill plateaus of Kandhamal district of Orissa (India) were analyzed to find out the monthly climatic index from the calculated values of effective rainfall and evapotranspiration. The 80% dependable monthly climatic index was correlated with crop coefficient and suitable cropping period and sequences for the study area were suggested based on it. The extent of investment, net return and soil loss from agriculture were estimated as per the present condition and for the suggested cropping patterns. A mathematical model was formulated for optimal allocation of area to different crop sequences with different objectives viz. minimization of soil loss, minimization of investment and maximization of net return from agriculture and was solved using linear goal programming technique. The model suggested to take up food crops in area of 1,30,777 ha and perennial grass cover in 3223 ha with a cropping intensity of 1.61 resulting in a net return of Rs 1064.775 millions sustaining soil loss to a tune of 9489.67 thousand tons per year. The model was found to be favourable in respect of higher net return of Rs 8862.34 and lesser soil loss of 23.41 tons/ha than the corresponding present values. But more investment of Rs 4489.01 per ha was required to fulfill the objectives.     

Bahiagrass crop coefficients from eddy correlation measurements in central Florida

Bahiagrass (Paspalum notatum) is a warm-season grass used primarily in pastures and along highways and other low maintenance public areas in Florida. It is also used in landscapes to some extent because of its drought tolerance. Bahiagrass can survive under a range of moisture conditions from no irrigation to very wet conditions. Its well-watered consumptive use has not been reported previously. In this study, bahiagrass crop coefficients (K _c) for an irrigated pasture were determined for July 2003 through December 2006 in central Florida. The eddy correlation method was used to estimate crop evapotranspiration (ET_c) rates. The standardized reference evapotranspiration (ET_o) equation (ASCE-EWRI standardization of reference evapotranspiration task committee report, 2005) was applied to calculate ET_o values using on site weather data. Daily K _c values were estimated from the ratio of the measured ET_c and the calculated ET_o. The recommended K _c values for bahiagrass are 0.35 for January–February, 0.55 for March, 0.80 for April, 0.90 for May, 0.75 for June, 0.70 for July–August, 0.75 for September, 0.70 for October, 0.60 for November, and 0.45 for December in central Florida. The highest K _c value of 0.9 in May corresponded with maximum vapor pressure deficit conditions as well as cloud free conditions and the highest incoming solar radiation as compared to the rest of the year. During the summer (June to August), frequent precipitation events increased the cloud cover and reduced grass water use. The K _c annual trend was similar to estimated K _c values from another well-watered warm-season grass study in Florida.