**3. Genetic assessment of sago palm**

*Genetic Variation*

**Figure 1.**

**Figure 2.**

*Morphological performance of sago palm forest.*

**32**

isolation [36].

for spiny formation in sago.

*with green color of the young leaf (D).*

by certain genes, so that there are certain nucleotide sequences in the sago palm genome that undergo transcription and translation processes in the spine formation process. Molecular markers encoding the characteristics of sago palm can be designed if desired by reverse transcription of sequences encoding protein synthesis

*Morphological performance of sago palm in the russet growth. Spineless with purple color of the young leaf (A), spineless with green color of the young leaf (B), spiny with purple color of the young leaf (C), and spiny* 

Genetic relatedness based on the phylogenetic constructs of each tested molecular marker shows that the distribution of the level of sample similarity according to the size of genetic distance is not limited by location and geographical isolation because samples from one island to others islands blending with each other [17]. The blending of Stylosanthes sp. obsessions from various regions in the dendrogram construction indicates that the obsessions have geographic distribution [46]. It was reported that, if there is distance and geographic isolation in the long term, the population from one region to another will experience differentiation or adaptive evolution as happened in Brassicaceae [47]. Furthermore, Scientist documented that population differentiation of O. rifipogon affected by distance or geographic

#### **3.1 Random amplified polymorphism DNA (RAPD) marker**

RAPD polymorphisms amplified on the PCR machine produced polymorphic fragments and the number of genotypes of each population. RAPD polymorphisms and high number of genotypes are a reflection of plant genetic diversity and adaptive evolution based on RAPD markers [38]. This result is in line with the diversities of sago palm revealed by using RAPD markers on several samples from Indonesia and Malaysia [48].

Population genetic diversity shows that the population samples from Papua have the highest of polymorphic sites number (S), the moderate of pairwise differences values (π), and the highest percentage of haplotype polymorphic compared to other populations from several islands in Indonesia [38]. Genotype diversity equal to one means that no identical genotype is found in a population sample. The value of Ĥ of individual samples at the island level all shows number of one, which means that one sample of individuals with another sample of individuals differs from one another based on the RAPD markers. Sago progenies obtained from semi-cultivated sago populations showed genetic differences among the progeny tested [43]. The varying values of S, π, and Ĥ indicate the genetic variation values of sago palm population in Indonesia. In the previous studies of sago palm by using RAPD markers showed that the sago palm diversities among individual was recorded also high [48] and other scientist reported 15 varieties of sago palm based on morphological characters in around Sentani lake [21].

The genetic hierarchy was estimated based on Analyses Molecular of Variance (AMOVA) calculations. The AMOVA calculation value shows that 89.35% of the total variety of samples is contributed by individuals with very significantly different with probability (P) values, 6.58% and 8.4% variance is contributed among populations [17]. The rates of diversities and adaptive evolution were detected in sago palm, those related to the genetic diversity of *M. sativa* L. by using RAPDs marker [47] as well as *Cynara scolymus* L. [49]. The statistical test method used to reveal the differentiation that occurs at the population and island level is also found to be used to reveal the differentiation that occurs at the population level in various types of plants, such as in *M. sativa* L [47], in *Acacia radiana* [50]*,* and in *Primula elatior* (L.) Oxlip [51].

Genetic relatedness among individual shows that the sago palm were classified into three groups based on the dendrogram construction. Group I include sago palm from all the populations as well as group III, while Group II includes sago palm from Jayapura, Serui, Manokwari and Ambon. This is related to the grouping of sago palm that it was reported in the previously study and divided sago palm from Indonesia and Malaysia into two groups and subgroups based on RAPD markers [48]. Previous sago genetic studies that focused on the Indonesian archipelago showed that sample individuals were divided into four groups based on RAPD markers [52]. Grouping of individuals in a dendrogram is largely determined by the genetic distance used, the method of grouping, and the desired bootstrap coefficient or rate. The differences between the groupings based on the cpDNA markers and the RAPD markers observed in previous studies are common in genetic relatedness studies [24, 39, 41].

Genetic relatedness among population shows a clustering pattern similar among individual. Genetic relatedness based on the dendrogram sample construction at the island level shows that the samples from the island of Papua are more closely related to the samples from Sumatra and Kalimantan, the samples from the island of Sulawesi are closely related to the samples from Ambon, and the samples from

the island of Java are separate from other islands based on the RAPD marker [38]. Here there is something interesting to observe because the sample at the island level forms a group together with samples from other islands that are far away, such as the sample from the island of Papua which forms a group together with the sample from the island of Sumatra. When examined from the migration side, it is possible that individual sago palm from Papua population have mingled with sago palm from the island of Sumatra. This phenomenon is possible because the molecular markers (RAPD) used are not as conservative as the cpDNA molecular markers that are uni-parental inherited [24, 25]. The RAPD marker is a nucleus genomic molecular marker associated with the DNA recombination process and is biparental inherited [39] so that the RAPD marker is a molecular marker that has a relatively short conservative time (one generation) compared to the cpDNA molecular marker. Previous studies suggest that higher variation is found using nucleus genomic markers rather than cpDNA markers [39–41, 53].
