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Dormancy and Germination of <I>Firmiana

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Dormancy and Germination of <I>Firmiana Luo, Zhang, Ren, Ma and Liu (2019). Seed Science and Technology, 47, 3, 343-349. https://doi.org/10.15258/sst.2019.47.3.09 Research Note Dormancy and germination of Firmiana danxiaensis, an endangered tree endemic to South China Xiaoying Luo1, Q ianmei Zhang2*, Hai Ren2*, Guohua M a2 and Hong L i u 3, 4 1 College of Tourism and Geography, Shaoguan University, Shaoguan 512005, China 2 Guangdong Provincial Key Laboratory of Applied Botany, South China Botanical Garden, Chinese Academy of Sciences, Guangzhou 510650, China 3 Department of Earth and Environment and International Center for Tropical Botany, Florida International University, Miami, FL 33199, USA 4 Forestry College, Guangxi University, Nanning 530004, China *Authors for correspondence (E-mail: [email protected]; [email protected]) (Submitted July 2019; Accepted October 2019; Published online November 2019) Abstract Firmiana danxiaensis (Malvaceae) is a rare and endemic tree of South China. It has a very narrow distribution with only two extant populations in the wild. The aim of this study was to understand the nature of its seed dormancy, how to break the dormancy and the environmental requirements for the natural germination of its seeds. To identify whether seeds of F. danxiaensis are dormant, germination experiments with 12 treatments (three light levels × four temperatures) were conducted. To determine the optimal way to break dormancy, we conducted the dormancy-breaking experiment with five treatments, including physical and chemical methods. The results showed that the dormancy of F. danxiaensis seeds was associated with a water- impermeable seed coat, i.e., the seeds have physical dormancy. Soaking the seeds in 98% concentrated sul phuric acid for one hour overcame the physical dormancy and enabled imbibition followed by germination (up to 73%). Once dormancy was broken, F. danxiaensis seeds could germinate over a wide range of temperatures in both light and darkness. The results suggest that ex situ conservation and reintroduction of F. danxiaensis might be achieved by sowing acid-treated seeds at restoration sites at the appropriate time of year. Keywords: acid scarification, endangered plant, Firmiana danxiaensis, germination, physical dormancy, South China Experimental and discussion The perennial deciduous tree Firmiana danxiaensis H.H. Hsue & H.S. Kiu (Malvaceae) was first reported in 1987 (Hsue et al., 1987). It was listed as a Second Class Key Protected Wild Plant of China in 1999, a Critically Endangered (CR) species in the China © 2019 Luo et al. This is an open access article distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: https://creativecommons.org/licenses/by-nc/4.0 343 XIAOYING LUO, Q IANMEI ZHANG, HAI REN, GUOHUA M A AND HONG L I U Species Red List in 2004, and a Plant with Extremely Small Populations (PESP) in China in 2012 (Ren et al., 2016). Firmiana danxiaensis has an extremely narrow distribution with only two known extant populations, which are located in the Danxia Landform in northern Guangdong Province in South China (Flora of China Editorial Committee, 2007). Research on F. danxiaensis has included tax onomy (Hsue et al., 1987; Qiu, 1994), popula tion genetics (Fan et al., 2013; Chen et al., 2014, 2015), floristic characteristics (Luo et al., 2015) and reintroduction trials (Zhang et al., 2014). Germination studies, however, have not been reported for F. danxiaensis, and little is known about the seed biology of this rare species. Seed ecology is important for the conservation of endangered plants (Herranz et al., 2010). Insufficient understanding of F. danxiaensis seed ecology hampers efforts to increase the population size of this plant through seed propagation. The current research aimed to identify reliable germination protocols to make ex situ propagation possible, which in turn would contribute to the species recovery program, including reintroduction, ex situ conservation and sustainable utilisation. Of the two extant populations of F. danxiaensis, one is located in the Danxiashan National Nature Reserve (113°36'25' - 113°47'53''E, 24°51'48'' - 25°04'12"N) (DX) and the other i s located in Cangshizai County Na ture Reserve (114°2'45"E, 25°08'19"N) (NX). Both locations are in Shaoguan City, northern Guangdong Province, South China. Mature seeds of F. danxiaensi s were manually collected from dehisced fruits at DX and NX on 20 August 2016. The seeds were then transferred to the laboratory, where they were stored in sealed plastic bags at ambient room temperature (25-30°C) and relative humidity (50-60%) for about one month until they were used in experiments. Seed viability was determined by using the 1% tetrazolium chloride (TTC) test on three repl icates of 15 decoated seeds at 30°C for 12 hours in darkness. Seeds with more than two-thirds of the embryo stained red were considered viable (Wu et al., 2016). The results showed that the seed viability was 58.3% for NX seeds and 81.8% for DX seeds. Water imbibition was assessed for (i) scarified see ds (a small hole was pierced in the seed coat at the chalazal end with a dissecting needle); (ii) decoated seed s (the entire seed coat was removed); and (iii) intact seeds. Three replica tes of 15 seeds were used for each treatment and population. The amount of water taken up was determined as the percentage increase in seed weight (Baskin et al., 2004): WI (%) = [(Wi – Wd) / Wd] × 100, where WI is percentage of increase in seed weights, Wi and Wd are the masses of imbibed and dry seeds, respectively. The imbibition curves of DX and NX seeds were very similar (figure 1). Intact seeds had only small increases (about 20%) in mass at 96 hours. When seeds were scarified, water absorption significantly increased (P < 0.01), i.e., the mass increased by 100% for both seed samples at 24 hours and increased to 120% for DX seeds and to 140% for NX seeds at 96 hours. In addition, the coat of some scarified seeds began to crack at 48 hours. The mass of decoated seeds increased to 130% for DX seeds and to 170% for NX seeds at 48 hours. To determine whether seeds of F. danxiaensis are dormant, we conducted a germina- tion experiment. The experiment included 12 treatments, i.e., three light levels × four temperatures. The three light levels were continuou s light, continuous darkness and 12 hours light / 12 hours dark. The four temperature treatments were a constant 25°C and three 1 2 hours light / 12 hours dark daily alternating te mperature regimes of 35/25, 25/15 344 DORMANCY AND GERMINATION OF FIRMIANA DANXIAENSIS 180 DX 160 140 a b 120 100 (%) I 80 W 60 40 c 20 0 0 102030405060708090100 Time (hours) intact scarified decoated 180 NX a 160 140 b 120 100 (%) I 80 W 60 40 c 20 0 0 102030405060708090100 Time (hours) intact scarified decoated Figure 1. Percentage increase in weight of intact, scarified and decoated seeds of Firmiana danxiaensis in the imbibition experiment. Different letters indicate significant differences between treatments based on maximum mass increase (P < 0.01). DX = seeds collected from Danxiashan National Nature Reserve; NX = seeds collected from Cangshizai County Nature Reserve. a nd 20/10°C. Three replicat es of 15 seeds were used for each treatment. The experiment was terminated after four weeks. The final germination percentage (GP), germination index (GI) and germination rate index (GRI) were calculated. GI = ∑(Gt / Dt), where Gt -1 is the number of germinated seeds after t days (Dt) (Shen et al., 2016); GRI (% d ) = ∑[( Gi - Gi-1) / i], where i is the germination count-day, Gi is the percentage of seeds that germinated at time i and Gi-1 is the percentage of seeds that germinated the previous count-day (Cook et al., 2008). The germination of fresh DX and NX seeds ranged from 0.0 to 4.4% at three incubation temperatures (35/25, 25, 25/15°C). No seeds germinated at 20/10°C. The germination percentage was significantly lower than the seed viability percentage, which was 58.3% 345 XIAOYING LUO, Q IANMEI ZHANG, HAI REN, GUOHUA M A AND HONG L I U for NX seeds and 81.8% for DX seeds. Illumination did not affect germination. Freshly matured, viable seeds may be defined as dormant if no seeds or only a few seeds have germinated at a range of temperature regimes in both light/dark and continuous darkness after about four weeks (Baskin and Baskin, 2014). Based on this criterion, we conclude that freshly matured seeds of F. danxiaensis are dormant, i.e., the germination of fresh seeds was < 5% at 15-35°C with or without light. Embryo type was determined by excising and examining embryos from fully imbibed seeds. The determination of dormancy class for F. danxiaensis seeds was facilitated by the use of a dichotomous seed dormancy key (Baskin and Baskin, 2014). We conclude that F. danxiaensis seeds have physical dormancy (PY) based on the following results: 1) embryos extracted from imbibed seeds had differentiated organs and were fully developed; 2) imbibition by intact seeds was low, indicating the presence of a water-impermeable seed coat; and 3) scarified seeds became fully imbibed in 24 hours and germinated within four weeks. PY has been reported for F. simplex (L.) W. Wight (Wang, 1989), Apeiba tibourbou Aubl. (Daws et al., 2006) and Kosteletzkya virginica (L.) C. Presl ex A. Gray (Poljakoff-Mayber et al., 1994), which are the other members of the Malvaceae; the latter is one of 18 angiosperm families reported to have seeds with PY (Gama-Arachchige et al., 2013; Mahadevan and Jayasuriya, 2013).
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