Flow cytometric analysis of nuclear DNA content,mitotic chromosome number and protein separation by SDS-PAGE in three accessions of African locust bean (<Emphasis Type="Italic">Parkia biglobosa</Emphasis> Benth.)

Nuclear DNA of three accessions of Parkia biglobosa collected from three locations in northern Cross River State was investigated using a Patec PA II flow cytometer equipped with an argon ion laser (488 nm), and pictures of mitotic chromosomes were taken using a digital micro-camera (Canon) placed on the eye piece of a binocular microscope at 100X oil immersion. Metaphase chromosome counts of 2n = 22 for accessions A and C and 2n = 24 for accession B, were obtained and through flow cytometry, the three accessions were confirmed to be diploids. The nuclear DNA content and genome size for the accessions were 1.5085, 1.489, and 1.5266 pg (737.7054, 728.121, and 746.5074 Mbp) for accessions A, B, and C, respectively. In another experiment, leaf samples from greenhouse-germinated seeds were analyzed for variation in the banding pattern of the protein by SDS-PAGE in the three accessions. Protein was resolved into three banding groups according to their electrophoretic mobility: slow, medium, and fast, clustering between 100–200, 40–70, and 10–25 kDa, respectively. There was 76% similarity in the banding pattern between the accessions.     

Estimating canopy cover from color digital camera image of rice field

Canopy cover (CC) is a good predictor variable for plant growth parameters such as leaf area index and aboveground biomass. A nondestructive, low-cost, and convenient method is presented for estimating CC using digital camera image analysis. CC was estimated by the ratio of plant pixels to total pixels of digital camera image of rice field. To determine the criteria for segmenting the rice plant from variable soil background, three mosaic images for rice plant, flooded/bare soil, and algae-infested background were prepared from digital camera images that were taken in various field conditions. An image analysis program was developed in Visual Basic to extract red, green, and blue (RGB) features from the mosaic images, calculate RGB-based color indices, and compute the minimum segmentation error for separating rice plant from background. When judged by the segmentation error, modified excessive green index (MEGI) showed the highest potential for segmenting rice plant from flooded/bare soil background, followed by normalized green (g) and excessive green index (EGI). At the threshold MEGI value of 0.03, the segmentation error was the lowest as 0.13%. Any single index considered was not satisfactory in segmenting rice plant from algae-infested background. However, a discriminant function of 1.2553EGI + 0.01735G − 0.01474B was successful in segmenting rice plant from flooded/bare soil and algaeinfested background with segmentation errors of 0.34 and 1.17%, respectively. CC for four rice varieties from tillering to booting stage was estimated based on the threshold value of MEGI and discriminant function and also manually using commercial software. Both estimates of CC showed good relationship of r² = 0.94, suggesting that a digital camera could be used efficiently for measuring the CC of rice field.     

A network of superconducting gravimeters detects submicrogal coseismic gravity changes

With high-resolution continuous gravity recordings from a regional network of superconducting gravimeters, we have detected permanent changes in gravity acceleration associated with a recent large earthquake. Detected changes in gravity acceleration are smaller than 10(-8) meters seconds(-2) (1 micro-Galileo, about 10(-9) times the surface gravity acceleration) and agree with theoretical values calculated from a dislocation model. Superconducting gravimetry can contribute to the studies of secular gravity changes associated with tectonic processes.     

Noxa, a BH3-only member of the Bcl-2 family and candidate mediator of p53-induced apoptosis

A critical function of tumor suppressor p53 is the induction of apoptosis in cells exposed to noxious stresses. We report a previously unidentified pro-apoptotic gene, Noxa. Expression of Noxa induction in primary mouse cells exposed to x-ray irradiation was dependent on p53. Noxa encodes a Bcl-2 homology 3 (BH3)-only member of the Bcl-2 family of proteins; this member contains the BH3 region but not other BH domains. When ectopically expressed, Noxa underwent BH3 motif-dependent localization to mitochondria and interacted with anti-apoptotic Bcl-2 family members, resulting in the activation of caspase-9. We also demonstrate that blocking the endogenous Noxa induction results in the suppression of apoptosis. Noxa may thus represent a mediator of p53-dependent apoptosis.     

Development of mulberry varieties for sustainable growth and leaf yield in temperate and subtropical regions of India

Sericulture is an agro-based industry, which heavily depends on mulberry (Morus spp.) for its sustainability, as mulberry leaf is the only available feed for the silkworm Bombyx mori L. Sericulture, in West Bengal and other subtropical regions in India, suffers from lack of adequate quantity of quality mulberry leaf during the colder months, the best season for rearing the high yielding bivoltine silkworm hybrids. In order to develop mulberry varieties, which can sustain normal growth during these colder months to yield better, efforts have been made in different research institutes in India. The present report is part of such an effort in which accessions in germplasm bank were screened for better growth, leaf retention and leaf yield contributing associated traits. Twenty three selected parents were crossed, 3,500 seeds were sown, 2,700 seedlings were transplanted to progeny row trail, selected 210 hybrids were evaluated under primary yield trial and finally nine hybrids along with a control variety were studied in detail for all leaf yield contributing traits under final yield trial. Stability analysis was adopted to identify hybrids, which can yield stably across seasons. The hybrids CT-44 and CT-11 out yielding the control by 17.17 and 7.11% were selected as these hybrids yield 7.93 and 8.15 mt/ha leaf respectively during the colder months (February) for their direct use as cultivars in West Bengal and other subtropical areas of India to sustain bivoltine sericulture to produce quality and gradable silk fibers.     

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.