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The methods described in this chapter were developed to avoid toxic organic phase separation utilized in many low-cost DNA extraction protocols such as the CTAB method. The steps involve: (1) lysis of the plant material, (2) binding of DNA to silica powder under chaotropic conditions, (3) washing the bound DNA, and (4) elution of DNA from the silica powder. This method has been tested in several plant species and the applicability of such DNA preparations for molecular marker studies in barley is shown in Chap. 8.
A half diallel mating design was used to study the inheritance of anthracnose and turcicum leaf blight (TLB) in six sorghum cultivars. Applying pathogens inoculum separately and applying both pathogens simultaneously differently affected the reaction of each genotype. GA06/18 was resistant to both pathogens. GA06/106 x Epuripuri and MUC007/009 x Epuripuri showed high heterosis for resistance to both diseases indicating that they were good materials for sorghum breeding. Additive and non‐additive (dominance) variance components were almost equally reflected by equal contribution of both variances towards the anthracnose resistance suggesting that both additive and dominance gene effects were involved in anthracnose resistance. Contribution of additive gene effects towards TLB resistance was greater than non‐additive gene effects suggesting that additive gene effects were more important in controlling TLB resistance. This study highlighted that deployment of resistant varieties is the most cost effective way to manage both diseases especially when integrated with appropriate agronomy practices.
Of importance to the successful extraction of genomic DNA from plant tissues is the collection of the suitable material and proper storage of the tissues before DNA isolation. If the samples are not properly treated, DNA can be degraded prior to isolation. The rate of sample degradation can vary dramatically from species to species depending on the method of sample collection. Mechanisms of genomic DNA degradation include exposure to endogenous nucleases due to organellar and cellular lysis. To prevent this from occurring, leaf or root tissues are commonly flash frozen in liquid nitrogen and then stored at À80 C. At these temperatures, nucleases remain inactive and DNA is stable. Thawing of tissue in some species can lead to rapid degradation. Therefore, during the extraction procedure, it may be necessary to grind the tissue to a fine powder in the presence of liquid nitrogen and expose frozen tissue immediately to a lysis buffer containing EDTA, which inhibits nuclease activity. This chapter provides an alternative method for sample collection and storage. Silica gel is used to desiccate tissues at room temperature. This avoids the use of liquid nitrogen and storage at À80 C.
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