ALBERT

Notes: Sixteen populations of the wild mulberry, Morus serrata Roxb., were analysed for their genetic diversity with the aim of using them in introgressive breeding programmes with cultivated relatives. Five genets from each population were collected from different natural populations of M. serrata present in Uttaranchal and Himachal Pradesh in India, and diversity of morpho-anatomical traits and inter-simple sequence repeat (ISSR) markers were studied. Significant amounts of genetic diversity were observed among these populations for morpho-anatomical as well as DNA markers. The 17 ISSR primers generated a total of 95 DNA markers, 51 of which were polymorphic, revealing 67% polymorphism among the populations. The pair-wise genetic distance, estimated from these DNA markers varied from 0.091 between Urgam-3 and Kathpuria to 0.258 between Dakrakao-1 and Dunda with an average genetic distance of 0.165. Clustering analysis grouped these 16 populations into three broad groups. The grouping showed a moderate correlation with the geographical distances. Based on the morphological traits and molecular genetic variability, plants of Urgam-1, Bhowali farm, Nainitikar, Dunda or Korwa-2 can be selected for breeding and conservation programmes.

Notes: Helminthosporium leaf blight (HLB) is the most important disease constraint to wheat (Triticum aestivum L.) cultivation in the eastern Gangetic Plains of South Asia. A Helminthosporium Monitoring Nursery (HMN) including potential adapted and exotic sources of HLB resistance was developed in Bangladesh, India and Nepal to assess the stability of genetic resistance across locations. The 8th, 9th and 10th HMN assessed the HLB resistance and agronomic traits of 17 wheat genotypes across 20 environments of Bangladesh, India and Nepal in the 1999-2000, 2000-2001 and 2001-2002 cropping seasons, respectively. The area under the disease progress curve (AUDPC) for HLB, grain yield, thousand-kernel weight (TKW), days to heading, days to maturity, and plant height were examined. The 17 genotypes showed a range of variability for disease and agronomic characters. Disease severity (AUDPC) differed in the 3 years and showed the highest values in 2002. The increase in AUDPC in 2002 caused the lowest grain yield, with an average 18% reduction due to HLB. A few genotypes (SW 89-5422, Yangmai-6 and Ning 8201) appeared to have stable HLB resistance across environments. However, most of the higher-yielding genotypes, except BL 1883, were unstable. The results suggest that careful selection of HLB resistance with acceptable grain yield, TKW and plant height may be possible using the wheat genotypes included in the HMN.

Notes: With a long-term plan to develop transgenic pigeonpea with resistance to fungal disease, the transfer of a rice chitinase gene to pigeonpea [Cajanus cajan (L.) Millsp.] is reported here. The rice chitinase gene harboured in the plasmid pCAMBIA 1302:RChit was delivered via the Agrobacterium-mediated method to the cotyledonary node explants followed by subsequent regeneration of complete plants on selection media containing hygromycin. Putative transformed pigeonpea plants were recovered with stringent selection pressure and confirmed using molecular techniques. Stable integration and expression of the chitinase gene has been confirmed in the T0 and T1 transgenics through molecular analysis.

Notes: The effect of abscisic acid (ABA) on the tolerance to oxidative stress in a freshwater green alga, Chlamydomonas reinhardtii, was investigated. Exogenously added ABA enhanced the growth of this alga, which was observed under continuous illumination but not in the dark. The cells treated with ABA for 24 h showed tolerance to oxidative stress caused by exposure to paraquat or hydrogen peroxide. In the ABA-treated cells, the activities of two antioxidant enzymes, catalase (CAT) and ascorbate peroxidase (APX), were significantly higher than those in the untreated control. The result suggests that ABA plays a role in the enhancement of tolerance to oxidative stress by increasing the activity of antioxidant enzymes.

Notes: The barley cultivar ‘Lenins’ was found to be a genotype showing high shoot regeneration ability in cultures derived from immature embryos. Five cultivars different from ‘Lenins’ in shoot regeneration ability were reciprocally crossed with ‘Lenins’ and the inheritance of tissue culture traits was investigated. F2 plants showed continuous distributions in callus growth and percentage of shoot regeneration, suggesting that these traits were controlled by polygenes. The F2 population, derived from a cross between ‘Lenins’ and ‘6721′, showed a monogenic segregation for the number of regenerated shoots, and the segregation ratio fitted 1:2:1. Tissue culture traits of ‘Lenins’ were controlled by several genes, whereas the number of regenerated shoots related to the efficiency of shoot regeneration is controlled by one major gene.

Notes: The aim was to develop an efficient chromosome doubling method for Miscanthus sinensis to enable the production of triploids and so avoid seed dispersal to the environment. Antimitotic treatments with colchicine or oryzalin were tested in M. sinensis cl. MS-88-110 on: (1) in vitro shoots and plants established in soil; (2) during propagation of embryogenic callus; and (c) during the initial stages of callus induction. All systems produced chromosome-doubled plants. A higher percentage of tetraploids was found after antimitotic treatment at the explant or callus level compared with treatment of in vitro shoots or plants established in soil. In general, oryzalin was more toxic to plant material than colchicine. A higher frequency of chimeras was found among plants with altered ploidy level when the target was formed shoot buds compared with adventitious shoot formation from callus. Antimitotic treatment of embryogenic callus from shoot apices also resulted in a high degree of albinism.

Notes: Effects of the resistant cultivars of melon on the powdery mildew fungus, Sphaerotheca fuliginea, were investigated in a melon-growing area of Japan. Powdery mildew fungi were isolated from four melon cultivars (‘HN21’, ‘Quincy’, ‘Earl's Knight Natsu 2’ and ‘Earl's Miyabi Natsu 2′) which are resistant to different races of S. fuliginea.‘HN21’is resistant to races 1 and N1,‘Quincy’ and ‘Earl's Knight Natsu 2’are resistant to races 1 and N2, and ‘Earl's Miyabi Natsu 2’is resistant to races 1, N1 and N2. All 155 strains isolated from ‘HN21’were identified as race N2. Seventy out of 81 strains isolated from ‘Quincy’ and ‘Earl's Knight Natsu 2’were identified as race N1, while other strains were identified as races 1, 2US, N2 and N3. Nineteen out of 29 strains isolated from ‘Earl's Miyabi Natsu 2’were identified as race N3, while other strains were identified as races 10, 2US, N1 and N4. These results suggest that, although the resistant cultivars are extremely effective for protecting the plants from their respective races, a race succession rapidly takes place, even in a small area, and other minor races become predominant.

Notes: Eighty-four cultivars of barley (Hordeum vulgare) and 95 wild strains (82 of H. spontaneum and 13 of H. agriocrithon) were surveyed for the production of callus, callus growth, and shoot regeneration in cultures derived from immature embryos. All cultivars except for ‘Turkey 381′, induced calli from more than 90% of embryos. On the other hand, the wild lines showed a large variation in the percentage of callus induction from 0 to 100%. Among the cultivars, those with the brittle rachis genotype, bt Bt2, on chromosome 3H, regenerated shoots with a significantly higher percentage than the cultivars with the Bt bt2 genotype. Green shoots were produced in a higher ratio (0.84) in the cultivars than in the wild lines (0.52). Among the lines examined,‘Lenins’ regenerated shoots efficiently (90.4%) and produced the highest number of calli with green shoots per embryo (4.77) followed by ‘Golden Promise’ (3.15). Examination of callus growth and shoot regeneration from embryos at different developmental stages revealed that scutellum development affected the quantity and quality of callus and shoot regeneration.

Notes: The present study, undertaken as a continuation of an earlier study on quantitative trait loci (QTL) analysis of grain protein content (GPC) in bread wheat (Prasad et al. 1999), includes the following: (1) identification of an additional molecular marker associated with GPC; (2) development of near-isogenic lines (NILs) for high GPC; and (3) the use of three sets of NILs (a total of 10 NILs) to validate the two available markers linked with QTL for GPC. A total of 114 sequence-tagged microsatellite site (STMS) primer pairs (that were not used in the previous study) were used for detection of polymorphism between the two parents (PH132, with high GPC; WL711, with low GPC) of a mapping population of 100 recombinant inbred lines (RILs). A total of 95 primer pairs gave amplification products, of which only 30 detected reproducible polymorphism between the parental genotypes. Bulked segregant analysis was conducted using these 30 primers on two bulks (each comprising eight RILs) representing the two extremes of the normal distribution. A solitary primer pair (WMC415) showed association with GPC, which was further confirmed through selective genotyping. Subsequently, 100 RILs were genotyped. A single-marker linear regression analysis showed significant association between the marker WMC415 and GPC, thus identifying a quantitative trait locus (designated as QGpcccsu-5A1), which explained 6.21% of the variation for GPC among the RILs. The above STMS marker, together with the STMS marker (WMC41) identified earlier, explains approximately 25% of the variation for GPC. In order to conduct validation of the above two available markers, 10 NILs were developed for high GPC using two genotypes (WL711 and HD2329) with low GPC as recipient parents and another two genotypes (PH132 and PH133) with high GPC as donor parents. NIL 2233 (with 11.7% GPC), derived from HD2329, when tried with WMC41 gave a characteristic amplification profile similar to that of its donor parent PH132, and NIL 2215 (with 11.9% GPC) derived from WL711, when tried with WMC415 gave an amplification profile that resembled its donor parent PH133. The remaining eight NILs with high GPC gave patterns similar to those of their corresponding recipient parents with both the markers, suggesting that either the QTL, other than those associated with the above markers, were actually transferred from the donor parents and contributed to high GPC in these NILs or that recombination had occurred between the markers identified and the corresponding QTL. Thus, the marker validation conducted using NILs, while demonstrating the utility of these two microsatellite markers for use in marker-assisted selection in plant breeding, also suggested that many more QTL exist that would need to be identified using closely linked molecular markers.