甜菜SSR反应体系优化及重要农艺性状分子标记

[目的]为选育优质甜菜新品种,利用SSR法筛选与高糖、高产、耐盐相关的分子标注.[方法]研究对甜菜SSR-PCR反应体系进行优化,并利用SSR分子标记方法和分离群体分组分析法(Bulked SegregateAnalysis,BSA)对具有高糖/低糖、低产/高产、耐盐/不耐盐三种重要农艺性状的甜菜亲本和F2代植株进行分析.[结果]研究建立了适宜甜菜的SSR-PCR反应体系为:20μL反应体系中含l×Buffer、2.0 mmol/LMg2+、1.5 uTaq DNA聚合酶、0.20 mmol/L dNTP、1.5 μmol/L引物、60 ng DNA模板.依据优化体系,对不同农艺性状的甜菜亲本与F2代进行SSR-PCR扩增分析,高糖性状获得了200和100 bp两条与高糖性状连锁的标记,250和230 bp两条与高产性状连锁的标记,550、250和100 bp三条与耐盐性状紧密连锁的分子标记.[结论]研究获得的7条特异条带是与甜菜重要农艺性状连锁的分子标记,将为甜菜的育种工作提供重要的理论基础.     

绵羊LDHβ基因的生物信息学分析及其在不同剩余采食量绵羊肝脏和肌肉组织中的表达

【目的】为鉴定绵羊乳酸脱氢酶β(lactate dehydrogenase B,LDHβ)基因的分子特征,研究其在不同剩余采食量绵羊肝脏和肌肉组织中的表达差异.【方法】测定了137只‘湖羊’公羔的剩余采食量(residual feed intake,RFI),按RFI进行排序,分别筛选出RFI最高(high-residual feed intake,H-RFI)和RFI最低(low-residual feed intake,L-RFI)的羊各15只,屠宰后,采集肝脏和肌肉组织,利用Q-PCR技术检测绵羊LDHβ基因分别在H-RFI和L-RFI羊肝脏和肌肉中表达量,并利用生物信息学软件构建了该基因的系统进化树和预测其结构与功能.【结果】绵羊LDHβ基因开放阅读框(open reading frame,ORF)为1 005bp,编码334个氨基酸,其蛋白质分子质量为36 479.43U,理论等电点为6.40.绵羊LDHβ基因与山羊的亲缘关系较近,其次为牛,序列同源性高.功能结构域预测结果显示,该基因编码产物在内质网中参与辅酶因子生物合成的可能性最高.Q-PCR结果显示,绵羊LDHβ-mRNA基因在L-RFI羊肝脏和肌肉中表达量均显著低于H-RFI羊(P0.01).【结论】绵羊LDHβ基因作为能量代谢的关键酶参与绵羊饲料效率的调控.     

Marker‐assisted introgression of rare allele of β‐carotene hydroxylase (crtRB1) gene into elite quality protein maize inbred for combining high lysine,tryptophan and provitamin A in maize

Vitamin A deficiency in humans is a widespread health problem. Quality protein maize (QPM) is a popular food rich in lysine and tryptophan, but poor in provitamin A (proA). Here, we report the improvement of an elite QPM inbred, HKI1128Q for proA using marker‐assisted introgression of crtRB1‐favourable allele. HKI1128 was one of the parental lines of three popular hybrids in India and was converted to QPM in our earlier programme. Severe segregation distortion for crtRB1 was observed in BC₁F₁, BC₂F₁ and BC₂F₂. Background selection by 100 SSRs revealed mean recovery of 91.07% recurrent parent genome varying from 88.78% to 93.88%. Across years, introgressed progenies possessed higher mean β‐carotene (BC: 9.22 µg/g), β‐cryptoxanthin (BCX: 3.05 µg/g) and provitamin A (proA: 10.75 µg/g) compared to HKI1128Q (BC: 2.26 µg/g, BCX: 2.26 µg/g and proA: 3.38 µg/g). High concentration of essential amino acids, viz. lysine (mean: 0.303%) and tryptophan (0.080%) in endosperm, was also retained. Multi‐year evaluation showed that introgressed progenies possessed similar grain yield (1,759–1,879 kg/ha) with HKI1128Q (1,778 kg/ha). Introgressed progenies with higher lysine, tryptophan and proA hold immense potential as donors and parents in developing biofortified hybrids.     

Concentration of selected heavy metals in spices,dry fruits and plant nuts

Different spices, dry fruits and plant nuts commonly consumed in Pakistan were assayed for the heavy metals cadmium, lead, copper, zinc, iron and manganese by the potentiometric stripping analysis and AA spectrophotometry. The results revealed wide variation in heavy metal content among different biological materials. Mixed spices generally exhibited higher value for trace metals specially lead (6.6–9.2 µg/g), cadmium (0.65–1.34 µg/g), iron (142.3–285.0 µg/g) and zinc (64.2–65.8 µg/g). Dry fruits contained relatively lesser amounts of heavy metals than plant nuts. Almonds contained higher levels of lead (1.02 µg/g) and cadmium (0.24 µg/g) than other nuts and dry fruits.     

Nutrient digestibility studies in Heteropneustes fossilis (Bloch), Clarias batrachus (Linnaeus) and C. gariepinus (Burchell)

The apparent digestibility coefficient (ADC %) for protein in catfishes, Clarias batrachus (Linnaeus) (16–18 cm; 45–50 g), C. gariepinus (Burchell) (21–22 cm; 60–65 g) and Heteropneustes fossilis (Bloch) (10–12 cm; 10–15 g) was estimated for nine feedstuffs of plant and animal origin. Fat digestibility was tested for C. batrachus and C. gariepinus, wherein the influence of feeding frequency on protein digestibility was also examined. The digestibility trials were conducted with triplicate groups of 25 fish each in 70‐L polyvinyl indoor flow‐through (1–1.5 L min^?1) circular tanks (water volume=55 L). Fish were fed to satiation daily, at 08:00 and 18:00 hours, and faecal matter was collected through slow siphoning. For studying the effect of feeding frequency, fish were fed the experimental diets at 08:00, 11:00, 14:00 and 17:00 hours. The protein digestibility of the feedstuffs varied significantly within each test species. However, the protein digestibility values for most of the ingredients tested were comparable between the three catfish species. The most efficient utilization of protein in these fishes was noted for soybean meal followed by lab‐prepared fishmeal. The lowest protein digestibility was recorded for rice bran. Similarly, the utilization of fat from the feedstuffs was also significantly different within each of the two species, C. batrachus and C. gariepinus. Fat digestibility of feed ingredients was similar between the two species. With respect to the fat, soybean meal, lab‐prepared fishmeal, full‐fat soybean meal and dried fish were more efficiently utilized, while the lowest fat digestibility was obtained for rice bran. A marked decline in protein utilization was observed in the two species where the feeding frequency was increased from two to four times a day.     

Diurnal variations in hypothalamic monoamine levels in the teleostChanna punctatus (bloch) in response to melatonin under two photothermal conditions

Hypothalamic dopamine (DA), noradrenaline (NA) and 5-hydroxytryptamine (5-HT) levels exhibited marked day-night variations under ambient photoperiod and temperature (12L12D; 17±1°C) with peak values at mid-light phase. The 16L8D; 22±1°C treatment reversed the diurnal rhythm of 5-HT, but not that of DA and NA. However, there was an overall increase in the levels of the catecholamines on exposure to the long photoperiod and high temperature. The administration of melatonin in the fish held on 16L8D; 22±1°C regime restored the 5-HT rhythm to that of the 12L12D; 17±1°C control group, but with elevated mid-photophase value. However, there was no effect of the indole treatment on the 5-HT rhythm under the 12L12D; 17±1°C regime. Melatonin causes a significant reduction of NA level in both the groups, while DA level did not change in either group.     

Studies on the ecology of actinomycetes in soil—VIII: Distribution and characteristics of acidophilic actinomycetes

Acidophilic actinomycetes, growing between pH 3.5 and 6.5, were isolated from 17 natural soils and mine wastes. Most isolates were Streptomyces spp. and selected ones were compared with some established neutrophilic species by assessment of overall similarity. Acidophilic and neutrophilic strains clustered separately, differing in a number of cultural and physiological characters, in addition to pH requirements. The possible importance of acidophilic actinomycetes in soil is discussed.     

Boron Induced Oxidative Stress,Antioxidant Defence Response and Changes in Artemisinin Content in Artemisia annua L.

Boron is an essential plant micronutrient and the range between deficient and toxic levels of boron is narrow for most of the plants. Like other elements, boron becomes toxic to growth at high concentrations. High boron concentrations in soil reduce crop productivity in many areas of the world. The effect of increasing levels of boron (0, 0.50, 1.00, 1.50, 2.00 mm ) on oxidative stress, antioxidant defence response and changes in artemisinin content in Artemisia annua were investigated in the present study. Boron toxicity reduced the growth parameters viz. stem height, fresh weight and dry weight. Treatments induced oxidative stress resulting in lower net photosynthetic rate, stomatal conductance, internal CO₂ and total chlorophyll content. The increased activities of antioxidant enzymes like CAT, POX and SOD were also noted in response to increasing levels of boron stress. However, H₂O₂ and artemisinin content were found to be high up to 1.00 mm concentration of boron compared to control, and on applying higher doses, further reduced contents were obtained. Thus, the results suggest that a mild stress of boron can be utilized for enhanced artemisinin production.