TITLE: No Title
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[DESCRIPTION]
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<p>The corms and leaves of elephant foot yams (<em>Amorphophallus paeoniifolius</em> (Dennst.) Nicolson) are important foods in the local diet in many Asian regions. The crop has high productivity and wide agroecological adaptation and exhibits suitability for the agroforestry system. Although the plant is assumed to reproduce via panmixia, a comprehensive study on the genetic background across regions to enhance wider consumer palatability is still lacking. Here, ten informative microsatellites were analyzed in 29 populations across regions in India, Indonesia and Thailand to understand the genetic diversity, population structure and distribution to improve breeding and conservation programs. The genetic diversity was high among and within regions. Some populations exhibited excess heterozygosity and bottlenecking. Pairwise <em>F</em><sub><em>ST</em></sub> indicated very high genetic differentiation across regions (<em>F</em><sub><em>ST</em></sub> = 0.274), and the Asian population was unlikely to be panmictic. Phylogenetic tree construction grouped the populations according to country of origin with the exception of the Medan population from Indonesia. The current gene flow was apparent within the regions but was restricted among the regions. The present study revealed that Indonesia and Thailand populations could be alternative centers of the gene pool, together with India. Consequently, regional action should be incorporated in genetic conservation and breeding efforts to develop new varieties with global acceptance.</p>
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[GUIDELINES]
<p><strong>Leaf collection</strong></p>
<p>Leaf of Amorphophallus paeoniifolius was collected by using dry method. </p>
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<p><strong>DNA extraction </strong></p>
<p>DNA was extracted using the modified CTAB method from the original protocol described by Lian et al<em>.</em> [2002]. Modification was required because the leaf of <em>A. paeoniifolius</em> contained glucomannan, a kind of high molecular of carbohydrate that difficult to dissolve in the water. Presence of glucomannan in the solution might cause DNA impurity, resulting on low PCR amplification. To separate DNA from the glucomannan, we used fishing method. Glucomannan pellet commonly transparent but still visible to be observed at the bottom of the extraction tube after we added water. After about 40 minutes, and DNA was expected dissolve completely, we pick up the glucomannan by fishing. For SSR analysis, DNA from approximately one gram of dry leaf was then preserved in a final volume of 200 µL of water and was stored at -30 °C until ready for use.</p>
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<p><strong>SSR primer optimation and reagent</strong></p>
<p>Polymerase chain reaction (PCR) was carried out in a 5-µL tube. For large sample, we recommend to use PCR plate with 48 wells. A 0.5-μL aliquot of extract was used in each PCR amplification to approximately contain 5-20 ng of DNA. SSR marker and DNA were amplified with a PCR reagent mix of 0.5 µM of the Forward primer, 0.1 µm of the Reverse primer tailed with U-19 (Fastac Co, Tokyo-Japan), 0.5 µM of the U-19 primer labeled with Texas Red, 0.2 mM of each dNTP, 1× PCR buffer (Mg<sup>2+</sup> free, Applied Biosystems), 2.5 mM MgCl<sub>2</sub>, and 0.5 U of Ampli Taq Gold DNA polymerase (Applied Biosystems). </p>
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