Maize Genomics and Genetics 2024, Vol.15, No.1, 27-35 http://cropscipublisher.com/index.php/mgg 28 strengths and limitations of this approach, also aims to identify gaps in the current research and suggest directions for future studies that could integrate isoenzymatic data with other phylogenetic tools to provide a more comprehensive understanding of Zea evolution. 1 Literature Review 1.1 Historical context of Zeaphylogenetic research Maize (Zea mays) is one of the world's most important crops. The study of its phylogenetic relationships is not only theoretically significant but also provides crucial information for breeding programs. As early as the early 20th century, scientists began to focus on the classification and phylogenetic relationships of maize and its wild relatives (García-Martínez and Martı́nez-Izquierdo, 2003). Through morphological, cytological, and molecular marker techniques, researchers gradually revealed the phylogenetic relationships among different species of maize. However, with the advancement of molecular biology technologies, especially genome and molecular marker techniques, research on the phylogenetic relationships of maize has entered a new phase. 1.2 Isoenzymatic variation in plants Isoenzymes, as a type of polymorphic marker, have been widely used in plant genetic diversity and phylogenetic studies. Research on isoenzymatic variation began in the 1960s when researchers separated and identified isoenzymes in different plant populations using electrophoresis (Matsuoka et al., 2002). Since then, numerous studies have found that isoenzymatic variation not only reflects genetic differences between plant populations but also reveals their evolutionary history and phylogenetic relationships (René Hortelano et al., 2011). In maize and its wild relatives, significant progress has also been made in the study of isoenzymatic variation. Studies have found that different species of maize and its relatives exhibit clear polymorphisms in isoenzymes such as esterases and peroxidases. These polymorphisms can serve as effective markers for identifying populations and studying phylogenetic relationships (Galal et al., 2013). 1.3 Comparative methods used in phylogenetic studies Comparative methods are crucial for revealing phylogenetic relationships in phylogenetic studies. Common comparative methods include morphological comparison, molecular marker analysis, and genome comparison. In morphological comparison, researchers infer phylogenetic relationships by comparing the morphological characteristics of different plants, such as leaf shape and inflorescence (Orton et al., 2017). However, because morphological characteristics are easily influenced by environmental factors, this method has limitations in revealing deep phylogenetic relationships. Molecular marker analysis is one of the most commonly used methods in phylogenetic studies. By analyzing isoenzymes, polymorphic DNA (such as RAPD, SSR), and gene sequence variations, researchers can more accurately reveal genetic relationships and evolutionary history among plant populations (Tsanev et al., 2000). Additionally, genome comparison methods provide more comprehensive phylogenetic information by comparing the genome structures and sequences of different species (Silva et al., 2020). 1.4 Significance of isoenzymatic markers in genetic studies Isoenzymatic markers have significant importance in plant genetic studies. Isoenzymatic markers have high polymorphism, which can reflect genetic differences between populations (Moeller and Tiffin, 2005). Isoenzymatic markers are easy to operate, as they can be obtained through electrophoresis, making them widely applicable in large-scale population studies (Gómez-Anduro et al., 2011). Finally, isoenzymatic markers can reveal the evolutionary history and phylogenetic relationships of populations, providing important references for breeding programs (Hartings et al., 2002). In maize and its wild relatives, studies on isoenzymatic markers have not only revealed genetic differences between different populations but also their phylogenetic relationships. For example, studies have found that maize and teosinte (Zea mexicana) exhibit high similarity in isoenzymes, providing strong evidence for their phylogenetic relationship (Angelov, 2003).
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