Transposable elements have been utilized as mutagens to create mutant libraries for functional genomics. Isolation of genomic segments flanking the insertion Mutator （Mu） is a key step in insertion mutagenesis studi...Transposable elements have been utilized as mutagens to create mutant libraries for functional genomics. Isolation of genomic segments flanking the insertion Mutator （Mu） is a key step in insertion mutagenesis studies. Herein, we adopted a modified AFLP method to identify and isolate Mu-flanking fragments from maize. The method consists of the following steps： 1） double-digestion of genomic DNA with Bgl II/Msp I and ligation of digested fragments to the Bgl II- and Msp I-adaptors; 2） enrichment of a subset of Bgl II/Msp I fragments followed by selective amplification of the Mu-flanking fragments; 3） simultaneous display of AFLP bands derived from the flanking regions for both insert and native Mu transposons; 4） identification and isolation of AFLP bands resulting from Mu insertions by comparing the banding profiles between Mu-induced mutants and their parental lines; and 5） confirmation of flanking fragments related to these Mu insertions. Using this approach, we have isolated flanking fragment（s） resulting from Mu insertion for every Mu-induced mutant, and one such fragment, M196-FF, is found to contain a partial sequence of the DNA topoisomerase I gene Topl. Moreover, the modified AFLP method including all restriction enzymes, adaptors and primers has been optimized in this study. The modified AFLP method has been proved to be simple and efficient in the isolation of Mu-flanking fragments and will find its usefulness in the functional genomics of maize.展开更多
Application of nitrogen (N) fertilizer is one of the most important approaches on improving maize grain yield. However, as is known to all, overuse N fertilizer not only leads to decline of N use efficiency and maize ...Application of nitrogen (N) fertilizer is one of the most important approaches on improving maize grain yield. However, as is known to all, overuse N fertilizer not only leads to decline of N use efficiency and maize yield, but also leads to potential risk to environment pollution. This experiment was conducted to determine the effects of N fertilizer applications with nine different treatments on soil physical-chemical characters and maize grain yield using hybrid variety Zhengdan 958 in 2011 and 2012. Results indicated that the soil bulk densities of T2 (CK) and T1 were the lowest compared to other treatments in 2011 and 2012, respectively, whereas the soil bulk density of T5 in 2011 and T3 in 2012 were higher than other treatments. The soil porosity and field capacity of T5 in 2011 and T3 in 2012 were lower than other treatments, but those of CK in 2011 and T1 in 2012 were higher than other treatments. The pH values of T3 to T7 were lower than other treatments. These results indicated that the soil bulk densities were increased, whereas the soil porosity, field capacity and values pH were decreased by N application at different stages. N application could increase the N contents of leaf and stem, whereas less or excess N application should not significant improve maize yield. Although the soil organic matter and total N contents of T3 were the highest in both 2011 and 2012, the yield of T4 is the highest in both 2011 and 2012. The application amount, period and times of N fertilizer were important to maize yield.展开更多
文摘Transposable elements have been utilized as mutagens to create mutant libraries for functional genomics. Isolation of genomic segments flanking the insertion Mutator （Mu） is a key step in insertion mutagenesis studies. Herein, we adopted a modified AFLP method to identify and isolate Mu-flanking fragments from maize. The method consists of the following steps： 1） double-digestion of genomic DNA with Bgl II/Msp I and ligation of digested fragments to the Bgl II- and Msp I-adaptors; 2） enrichment of a subset of Bgl II/Msp I fragments followed by selective amplification of the Mu-flanking fragments; 3） simultaneous display of AFLP bands derived from the flanking regions for both insert and native Mu transposons; 4） identification and isolation of AFLP bands resulting from Mu insertions by comparing the banding profiles between Mu-induced mutants and their parental lines; and 5） confirmation of flanking fragments related to these Mu insertions. Using this approach, we have isolated flanking fragment（s） resulting from Mu insertion for every Mu-induced mutant, and one such fragment, M196-FF, is found to contain a partial sequence of the DNA topoisomerase I gene Topl. Moreover, the modified AFLP method including all restriction enzymes, adaptors and primers has been optimized in this study. The modified AFLP method has been proved to be simple and efficient in the isolation of Mu-flanking fragments and will find its usefulness in the functional genomics of maize.
文摘Application of nitrogen (N) fertilizer is one of the most important approaches on improving maize grain yield. However, as is known to all, overuse N fertilizer not only leads to decline of N use efficiency and maize yield, but also leads to potential risk to environment pollution. This experiment was conducted to determine the effects of N fertilizer applications with nine different treatments on soil physical-chemical characters and maize grain yield using hybrid variety Zhengdan 958 in 2011 and 2012. Results indicated that the soil bulk densities of T2 (CK) and T1 were the lowest compared to other treatments in 2011 and 2012, respectively, whereas the soil bulk density of T5 in 2011 and T3 in 2012 were higher than other treatments. The soil porosity and field capacity of T5 in 2011 and T3 in 2012 were lower than other treatments, but those of CK in 2011 and T1 in 2012 were higher than other treatments. The pH values of T3 to T7 were lower than other treatments. These results indicated that the soil bulk densities were increased, whereas the soil porosity, field capacity and values pH were decreased by N application at different stages. N application could increase the N contents of leaf and stem, whereas less or excess N application should not significant improve maize yield. Although the soil organic matter and total N contents of T3 were the highest in both 2011 and 2012, the yield of T4 is the highest in both 2011 and 2012. The application amount, period and times of N fertilizer were important to maize yield.
