Magnolia sirindhorniae Noot. & Chalermglin is an endangered species with high ornamental and commercial value that needs to be urgently protected and judiciously commercialized. In this study, a protocol for efficient regeneration of this species is standardized. The lateral buds of the M. sirindhorniae plant were used as an explant. Half-strength Murashige and Skoog (MS) medium supplemented with 2.0 mg/L 6-benzyladenine (BA), 0.1 mg/L α-naphthaleneacetic acid (NAA), and 2.0 mg/L gibberellic acid (GA 3 ) was found to be the optimal medium for shoot induction. The maximum shoot multiplication rate (310%) was obtained on Douglas-fir cotyledon revised medium (DCR) fortified with 0.2 mg/L BA, 0.01 mg/L NAA, and additives. The half-strength DCR medium supplemented with 0.5 mg/L NAA and 0.5 mg/L indole-3-butyric acid (IBA) supported the maximum rate (85.0%) of in vitro root induction. After a simple acclimatization process, the survival rate of plantlets in a substrate mixture of sterile perlite and peat soil (1:3; v/v ) was 90.2%. DNA markers were used for assessment of genetic uniformity, confirming the genetic uniformity and stability of regenerated plants of M. sirindhorniae . Thus, the described protocol can safely be applied for large scale propagation of this imperative plant.

Michelia shiluensis is a rare and endangered magnolia species found in South China. This species produces beautiful flowers and is thus widely used in landscape gardening. Additionally, its timber is also used for furniture production. As a result of low rates of natural reproduction and increasing levels of human impact, wild M. shiluensis populations have become fragmented. This species is now classified as endangered by the IUCN. In the present study, we characterized the complete chloroplast genome of M. shiluensis and found it to be 160,075 bp in length with two inverted repeat regions (26,587 bp each), a large single-copy region (88,105 bp), and a small copy region (18,796 bp). The genome contained 131 genes, including 86 protein-coding genes, 37 tRNAs, and 8 rRNAs. The guanine-cytosine content represented 39.26% of the overall genome. Comparative analysis revealed high similarity between the M. shiluensis chloroplast genome and those of four closely related species: Michelia odora, Magnolia laevifolia, Magnolia insignis, and Magnolia cathcartii. Phylogenetic analysis shows that M. shiluensis is most closely related to M. odora. The genomic information presented in this study is valuable for further classification, phylogenetic studies, and to support ongoing conservation efforts.

Toona ciliata (Meliaceae) is a rare, fast-growing commercial tree species that produces red timber with beautiful grains. It used to be widely distributed in the past, but few natural forests exist in the field now. Here, we tested 12 nuclear microsatellite markers derived from 551 individuals of 29 populations in China, and investigated the genetic diversity and population structure in T. ciliata. Our results revealed 146 alleles obtained from all of the populations studied here, with an average polymorphic information content of 0.723. The populations could be divided into two major genetic lineages using STRUTURE analysis: 14 and 15 populations from different provinces clustered in Western and Eastern genetic lineages, respectively. The genetic differentiation among the two genetic lineages was 28%. The genetic diversity of the Western lineage was higher than that of the Eastern lineage. Moreover, a Mantel test of the relationship between genetic and geographic distances revealed significantly positive correlations both among all 29 populations and within the Eastern genetic lineage, whereas there was no significant correlation in the Western genetic lineage. Therefore, in the conservation strategy of T. ciliatare sources, the two genetic lineages should be treated independently. To the best of our knowledge, these data are the first detailed characterization of the genetic structure and diversity of T. ciliata populations using nuclear microsatellite markers. The obtained results lay the foundation for preservation of the T. ciliata germplasm resources and support further artificial breeding efforts.

Research Highlights: This study is the first to examine the genetic diversity of Michelia shiluensis (Magnoliaceae). High genetic diversity and low differentiation were detected in this species. Based on these results, we discuss feasible protection measures to provide a basis for the conservation and utilization of M. shiluensis. Background and Objectives: Michelia shiluensis is distributed in Hainan and Guangdong province, China. Due to human disturbance, the population has decreased sharply, and there is thus an urgent need to evaluate genetic variation within this species in order to identify an optimal conservation strategy. Materials and Methods: In this study, we used eight nuclear single sequence repeat (nSSR) markers and two chloroplast DNA (cpDNA) markers to assess the genetic diversity, population structure, and dynamics of 78 samples collected from six populations. Results: The results showed that the average observed heterozygosity (Ho), expected heterozygosity (He), and percentage of polymorphic loci (PPL) from nSSR markers in each population of M. shiluensis were 0.686, 0.718, and 97.92%, respectively. For cpDNA markers, the overall haplotype diversity (Hd) was 0.674, and the nucleotide diversity was 0.220. Analysis of markers showed that the genetic variation between populations was much lower based on nSSR than on cpDNA (10.18% and 77.56%, respectively, based on an analysis of molecular variance (AMOVA)). Analysis of the population structure based on the two markers shows that one of the populations (DL) is very different from the other five. Conclusions: High genetic diversity and low population differentiation of M. shiluensis might be the result of rich ancestral genetic variation. The current decline in population may therefore be due to human disturbance rather than to inbreeding or genetic drift. Management and conservation strategies should focus on maintaining the genetic diversity in situ, and on the cultivation of seedlings ex-situ for transplanting back to their original habitat.

Magnolia lucida (Magnoliaceae) is classified as an endangered species by the International Union for Conservation of Nature. It has high commercial value owing to its attractive tree shape and flowers. We adopted an excellent genotype of M. lucida as the parent material and established a mini-cut orchard through grafting to provide trunk shoots explants over the long-term. Optimal sterilization was achieved using a combination of 75% ethanol for 30 s, one percent benzalkonium bromide for five minutes, and 0.1% mercuric chloride for five minutes. Modified Murashige and Skoog medium (ML) was the optimal medium for the growth of M. lucida. Addition of one mg/L of 6-benzyl adenine (BA) and 0.05 mg/L of α-naphthaleneacetic acid (NAA) to the medium increased the shoot induction rate to 95.56%, and the ML medium containing 0.4 mg/L BA and 0.04 mg/L NAA achieved the maximum multiplication rate (284.56%). Dark treatment for seven days, followed by continuous light treatment could better resolve the challenge of difficult rooting in M. lucida plants. Using random amplified polymorphic DNA and inter simple sequence repeat markers, we confirmed the genetic uniformity and stability of the regenerated plants. Our protocol should be helpful for the propagation and conservation of this endangered plant.