Plant Industries, Northern Territory Department of Primary Industry and Fisheries, GPO Box 3000, Darwin, NT 0801, Australia Present address: New South Wales Department of Primary Industries, Tamworth Agricultural Institute, 4 Marsden Park Rd, Tamworth, NSW 2340, Australia
Plant Industries, Northern Territory Department of Primary Industry and Fisheries, GPO Box 3000, Darwin, NT 0801, Australia.
AoB Plants. 2014 Dec 30;7:plu091. doi: 10.1093/aobpla/plu091.
Understanding the root distribution of trees by soil coring is time -: consuming as it requires the separation of roots from soil and classification of roots into particular size classes. This labour-intensive process can limit sample throughput and therefore sampling intensity. We investigated the use of quantitative polymerase chain reaction (qPCR) on soil DNA extractions to determine live fine root DNA density (RDD, mg DNA m(-2)) for mango (Mangifera indica) trees. The specificity of the qPCR was tested against DNA extracted from 10 mango cultivars and 14 weed species. All mango cultivars and no weeds were detected. Mango DNA was successfully quantified from control soil spiked with mango roots and weed species. The DNA yield of mango root sections stored in moist soil at 23-28 °C declined after 15 days to low concentrations as roots decayed, indicating that dead root materials in moist soil would not cause false-positive results. To separate large roots from samples, a root separation method for field samples was used to target the root fragments remaining in sieved (minimum 2 mm aperture) soil for RDD comparisons. Using this method we compared the seasonal RDD values of fine roots for five mango rootstock cultivars in a field trial. The mean cultivar DNA yields by depth from root fragments in the sieved soil samples had the strongest relationship (adjusted multiple R(2) = 0.9307, P < 0.001) with the dry matter (g m(-2)) of fine (diameter <0.64 mm) roots removed from the soil by sieving. This method provides a species-specific and rapid means of comparing the distribution and concentration of live fine roots of trees in orchards using soil samples up to 500 g.
通过土壤芯样了解树木的根系分布非常耗时,因为需要将根系从土壤中分离出来,并将根系分类为特定的大小类别。这种劳动密集型过程可能会限制样本通量,从而限制采样强度。我们研究了使用定量聚合酶链反应 (qPCR) 对土壤 DNA 提取物进行分析,以确定芒果 (Mangifera indica) 树的活细根 DNA 密度 (RDD,mg DNA m(-2))。qPCR 的特异性已针对从 10 个芒果品种和 14 种杂草物种提取的 DNA 进行了测试。所有芒果品种和杂草均被检测到。成功地从用芒果根和杂草物种污染的对照土壤中定量芒果 DNA。将芒果根段储存在 23-28°C 的潮湿土壤中,15 天后,随着根系腐烂,DNA 产量下降到低浓度,表明潮湿土壤中的死根材料不会导致假阳性结果。为了将大根与样品分离,使用了一种针对现场样品的根系分离方法,旨在针对筛出的(最小孔径 2 毫米)土壤中剩余的根系片段进行 RDD 比较。使用这种方法,我们比较了五个芒果砧木品种在田间试验中的细根季节性 RDD 值。筛出土壤样品中根系碎片的深度平均品种 DNA 产量与筛出土壤中细根(直径 <0.64 毫米)的干物质(g m(-2)) 之间具有最强的关系(调整后的多重 R(2) = 0.9307,P < 0.001)。该方法提供了一种使用土壤样品(高达 500 g)快速比较果园中树木活细根分布和浓度的物种特异性方法。