Seed Characteristics and Germination Behaviour of *Bauhinia malabarica* Roxb.

*Pazhayaveetil Kuttanpillai Chandrasekhara Pillai, Sanal Chalil Viswanath, Thoduvayil Karunakaran Hrideek and Aviyan Hari Jiji*

#### **Abstract**

Malabar *Bauhinia* (*Bauhinia malabarica*) is a native ornamental species belonging to the family Fabaceae, distributed throughout India in semievergreen and moist deciduous forests and in gardens. Information regarding seed characteristics and seed handling of the species is meagre. This study describes seed characteristics, germination behaviour and pretreatment for enhancing seed germination of *B. malabarica*. Treating the seeds with concentrated sulphuric acid for 30 min reduced germination duration up to 6 days and increased germination rate up to 100% against 22 days of germination duration and 10% germination in seeds without treatment. The results of this study are helpful for conservation and nursery practices of *B. malabarica*.

**Keywords:** maturity index, seed germination, germination value, pretreatment, sulphuric acid

#### **1. Introduction**

*Bauhinia malabarica* Roxb., commonly known as Malabar *Bauhinia*/mountain ebony, is a deciduous tree belonging to the family of Fabaceae, distributed all over the Indo-Malayan region. The species is found throughout Indian subcontinent and popularised in different vernacular names as Arampuli (Malayalam), Malayathi (Tamil), Basavana pada (Kannada), Amli (Hindi), etc. In native range, the species is distributed in the semievergreen and moist deciduous forests up to 600 m. It is used as an ornamental plant in homesteads and gardens of the native range.

Studies of the genus *Bauhinia* is very limited compared with other genus like *Caesalpinia*, *Cassia*, *Tamarindus*, etc., under the family Fabaceae. Primary documentation of the genus *Bauhinia* in India was conducted during the eighteenth century [1]. Neotypification of the species *Bauhinia malabarica* was recently done by the Central National Herbarium of Botanical Survey of India [2]. Micromorphological characteristics of selected species of the genus *Bauhinia* were studied earlier [3]. Variation in size and structure of selected species of *Bauhinia* was examined and recorded by different scholars [4, 5]. Analysis on mineral elements and nutritional and anti-nutritional contents in the seeds of *B. monandra* was conducted in an earlier study [6]. Phytochemical structure like polysaccharide and

proteins in seeds of *Bauhinia* was investigated earlier [7, 8]. Antibacterial nature of *B. acuminata* was reported in a previous study [9]. It was reported that seed oil of *Bauhinia* is a novel substance for the production of sophorolipids [10].

A few investigations were done on *B. malabarica* on foliar micromorphology, natural regeneration, chemical composition, phytochemical analysis, antimalarial activities and anti-oxidant potential [3, 11–15]. Studies related to seed germination in the genus *Bauhinia* was conducted only on selected species like *B. rufescens*, *B. thonningii*, etc. [16–18]. However, information on seed characteristics and germination parameters of *B. malabarica* is limited. The present study was done to fill the above-mentioned gaps.

#### **2. Materials and methods**

Periodical observations were carried out on mother plants of *B. malabarica* to identify optimum maturity index for seed collection. Mature pods (fruits) of *B. malabarica* were collected (March–April, 2016) from Peechi-Vazhani Wildlife Sanctuary (10°31′48″N; 76°20′50″E) in Thrissur District, Kerala, India. The pods were dried under sunlight (35–38°C) for 2 days, and seeds were extracted by splitting the pods. Extracted seeds were dried in shade, cleaned and stored in airtight containers. Seed characteristics such as morphology, moisture content and germination were evaluated. High constant temperature oven-dry method was used to determine seed moisture content [19]. The seeds were dried in hot-air oven for 1 h at 130°C. Seed moisture content (MC %) was estimated according to the formula of ISTA.

$$\text{distance content} \stackrel{\textstyle M}{\ (\text{MC }\%)} \text{ was estimated according to the formula of ISTA.}$$

$$\text{MC }\% = \frac{\text{fresh weight of seed - over } dry \text{ weight of seed}}{\text{fresh weight of seed}} \times 100\tag{1}$$

Seeds were subjected to different pre-sowing treatments to enhance seed germination and reduce the germination period. The following were the pretreatments applied for the study:

T1: control (no pre-sowing treatment).

T2: soaked in water for 24 h.

T3: soaked in water for 48 h.

T4: soaked in hot water for 2 min.

T5: soaked in hot water for 5 min.

T6: soaked in hot water for 2 min + soaked in water for 24 h.

T7: soaked in hot water for 5 min + soaked in water for 24 h.

T8: soaked in GA3 (500 ppm) for 2 h.

T9: soaked in GA3 (1000 ppm) for 2 h.

T10: acid treatment (conc. H2SO4) for 10 min.

T11: acid treatment (conc. H2SO4) for 20 min.

T12: acid treatment (conc. H2SO4) for 30 min.

Tap water (≈35°C) was used in T2 and T3 treatments, whereas in the treatments T4–T7, hot water (85°C) was used. Different concentrations of gibberellic acid (GA3)/gibberellin A3 (chemical formula: C19H22O6) were used in T8 and T9 treatments. Concentrated sulphuric acid (98%) was used in the treatments T10–T12.

Seeds (n = 100 in 4 replications) were sown in germination trays having a size of 25 × 20 × 5 cm filled with vermiculite and kept in germination room (30 ± 2°C and 90% RH) under laboratory condition. Randomised block design was adopted for the experiment. Data on seed germination were recorded starting from seed germination till culmination and computed germination-related parameters. Germination initial time (GIT), germination percentage (GP), germination duration (GD), mean germination time (MGT), mean daily germination (MDG),

**5**

**3. Results**

*Seed Characteristics and Germination Behaviour of* Bauhinia malabarica *Roxb.*

Germination-related parameters were determined as follows:

where Dg = first germination day and Ds = seed sowing day.

where G = no. of germinated seeds and T = no. of seeds sown.

of sowing and G = total no. of germinated seeds.

germination energy (GE), peak value (PV) and germination value (GV) were

Germination initial time (GIT) = Dg–Ds (2)

Germination percentage (GP) = (G/T) × 100 (3)

Germination duration (GD) = Gf–Gi (4)

Mean germination time (MGT) = (Gt × Dt)/G (5)

where Gt = no. of germinated seeds at day-t, Dt = no. of days at 't' from the day

Mean daily germination (MDG) = GP/Gd (6)

where GP = germination percentage and Gd = no. of days to complete

(Xn − Xn − 1)/Yn (7)

where Xn = no. of germinants on the nth counting date and Yn = no. of days

Germination value (GV) = PV × MDG (9)

Each trait was analysed using mean values under the various pretreatments. The variation on mean values between these treatments were performed through analysis of variance (ANOVA) done by statistical software SPSS version 22.

The study recorded 7092 ± 50 seeds per kilogram. Moisture content (MC %) of

no.of days required to the peak germination (8)

Germination energy (GE) = X1/Y1 + (X2 − X1)/Y2 + .……+

Peak value (PV) = Highest number of seeds germinated/

where PV = peak value and MDG = mean daily germination.

where Gf = final day of germination and Gi = initial day of germination.

*DOI: http://dx.doi.org/10.5772/intechopen.84970*

calculated [20–24].

germination.

from sowing to the nth count.

**2.1 Statistical analysis**

fresh seeds was 5.35%.

**3.1 Seed weight and moisture content**

*Seed Characteristics and Germination Behaviour of* Bauhinia malabarica *Roxb. DOI: http://dx.doi.org/10.5772/intechopen.84970*

germination energy (GE), peak value (PV) and germination value (GV) were calculated [20–24].

Germination-related parameters were determined as follows:

$$\text{Germination initial time (GIT)} = \text{Dg-Ds} \tag{2}$$

where Dg = first germination day and Ds = seed sowing day.

$$\text{Germination percentage (GP)} = \text{(G/T)} \times 100\tag{3}$$

where G = no. of germinated seeds and T = no. of seeds sown.

$$\text{Germination duration (GD)} = \text{Gf} - \text{Gi} \tag{4}$$

where Gf = final day of germination and Gi = initial day of germination.

$$\text{Mean generation time (MGT)} = \text{(Gt} \times \text{Dt)}/\text{G} \tag{5}$$

where Gt = no. of germinated seeds at day-t, Dt = no. of days at 't' from the day of sowing and G = total no. of germinated seeds.

$$\text{Mean daily generation (MDG)} = \text{GP/Gd} \tag{6}$$

where GP = germination percentage and Gd = no. of days to complete germination.

$$\begin{aligned} \text{Germination energy (GE)} &= \text{X1/Y1+ (X2-X1)/Y2+ } \text{.......++} \\ &\quad \text{(Xn - Xn - 1)/Yn} \end{aligned} \tag{7}$$

where Xn = no. of germinants on the nth counting date and Yn = no. of days from sowing to the nth count.

$$\begin{array}{l}\text{Peak value (PV)} = \text{Highest number of seeds germinates} / \\ \text{noof days required to the peak germination} \end{array} \tag{8}$$

$$\text{Germination value (GV)} = \text{PV} \times \text{MDG} \tag{9}$$

where PV = peak value and MDG = mean daily germination.

#### **2.1 Statistical analysis**

Each trait was analysed using mean values under the various pretreatments. The variation on mean values between these treatments were performed through analysis of variance (ANOVA) done by statistical software SPSS version 22.

#### **3. Results**

#### **3.1 Seed weight and moisture content**

The study recorded 7092 ± 50 seeds per kilogram. Moisture content (MC %) of fresh seeds was 5.35%.

*Horticultural Crops*

**2. Materials and methods**

applied for the study:

T1: control (no pre-sowing treatment).

T8: soaked in GA3 (500 ppm) for 2 h. T9: soaked in GA3 (1000 ppm) for 2 h.

T10: acid treatment (conc. H2SO4) for 10 min. T11: acid treatment (conc. H2SO4) for 20 min. T12: acid treatment (conc. H2SO4) for 30 min.

T6: soaked in hot water for 2 min + soaked in water for 24 h. T7: soaked in hot water for 5 min + soaked in water for 24 h.

T2: soaked in water for 24 h. T3: soaked in water for 48 h. T4: soaked in hot water for 2 min. T5: soaked in hot water for 5 min.

proteins in seeds of *Bauhinia* was investigated earlier [7, 8]. Antibacterial nature of *B. acuminata* was reported in a previous study [9]. It was reported that seed oil of

Periodical observations were carried out on mother plants of *B. malabarica* to identify optimum maturity index for seed collection. Mature pods (fruits) of *B. malabarica* were collected (March–April, 2016) from Peechi-Vazhani Wildlife Sanctuary (10°31′48″N; 76°20′50″E) in Thrissur District, Kerala, India. The pods were dried under sunlight (35–38°C) for 2 days, and seeds were extracted by splitting the pods. Extracted seeds were dried in shade, cleaned and stored in airtight containers. Seed characteristics such as morphology, moisture content and germination were evaluated. High constant temperature oven-dry method was used to determine seed moisture content [19]. The seeds were dried in hot-air oven for 1 h at 130°C. Seed moisture content (MC %) was estimated according to the formula of ISTA.

*MC*%= *fresh weight of seed* <sup>−</sup> *oven dry weight of seed* \_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_ *fresh weight of seed* <sup>×</sup> <sup>100</sup> (1)

Seeds were subjected to different pre-sowing treatments to enhance seed germination and reduce the germination period. The following were the pretreatments

Tap water (≈35°C) was used in T2 and T3 treatments, whereas in the treatments

T4–T7, hot water (85°C) was used. Different concentrations of gibberellic acid (GA3)/gibberellin A3 (chemical formula: C19H22O6) were used in T8 and T9 treatments. Concentrated sulphuric acid (98%) was used in the treatments T10–T12. Seeds (n = 100 in 4 replications) were sown in germination trays having a size of 25 × 20 × 5 cm filled with vermiculite and kept in germination room (30 ± 2°C and 90% RH) under laboratory condition. Randomised block design was adopted for the experiment. Data on seed germination were recorded starting from seed germination till culmination and computed germination-related parameters. Germination initial time (GIT), germination percentage (GP), germination duration (GD), mean germination time (MGT), mean daily germination (MDG),

A few investigations were done on *B. malabarica* on foliar micromorphology, natural regeneration, chemical composition, phytochemical analysis, antimalarial activities and anti-oxidant potential [3, 11–15]. Studies related to seed germination in the genus *Bauhinia* was conducted only on selected species like *B. rufescens*, *B. thonningii*, etc. [16–18]. However, information on seed characteristics and germination parameters of *B. malabarica* is limited. The present study was done to fill the above-mentioned gaps.

*Bauhinia* is a novel substance for the production of sophorolipids [10].

**4**

#### **3.2 Maturity index**

The optimum maturity indices for seed collection of *B. malabarica* were presented in **Table 1**. The maturity indices identified for determining optimum period for seed collection were colour of pods (yellowish-green turned to blackish-green), leaf number (minimum number of leaves), dehydration (pods become dehydrated) and hardness (pods and seeds become hardest).

#### **3.3 Pod/seed characteristics**

Colour, shape, size, type, weight, number of seeds per pod and per kg, type of germination, etc. are presented in **Table 2**.

#### **3.4 Seed germination**

**Figure 1** depicts germination pattern of seeds under various pretreatments. Seed germination among treatments was significantly different (**Table 3**).

Germination-related parameters under different pretreatments such as germination initial time, germination percentage, germination duration, mean germination time, mean daily germination, germination energy, peak value and germination value are given in **Table 4**.


#### **Table 1.**

*Pod/seed maturity indices of B. malabarica.*


**7**

*Seed Characteristics and Germination Behaviour of* Bauhinia malabarica *Roxb.*

**Germination Initial Time (GIT)**: Germination initial time was 2 days after sowing in the treatments T4, T10, T11 and T12, whereas it was 5 days after sowing in

**Source Sum of squares Degrees of freedom (df) Mean square F-value Sig.** Between groups 4332.775 11 393.889 29.804 0.005

*Seed germination pattern under different treatments. Note: T1, control; T2, soaked in water for 24 h; T3, soaked in water for 48 h; T4, soaked in hot water for 2 min; T5, soaked in hot water for 5 min; T6, soaked in hot water for 2 min + soaked in water for 24 h; T7, soaked in hot water for 5 min + soaked in water for 24 h; T8, soaked in gibberellic acid (GA3—500 ppm) for 2 h; T9, soaked in gibberellic acid (GA3—1000 ppm) for 2 h; T10, acid treatment (conc. H2SO4) for 10 min; T11, acid treatment (conc. H2SO4) for 20 min; T12, acid treatment (conc. H2SO4) for 30 min.*

**Germination Duration (GD)**: Seed germination started 2 days after sowing and completed in 28 days. The least GD noticed in treatment T12 (06 days) followed by

**Mean Germination Time (MGT)**: Mean germination time was more in T12 treatment (21.67) where the treatment gave 100% germination, whereas the least

**Mean Daily Germination (MDG)**: Mean daily germination was highest in the

**Germination Energy (GE)**: Germination energy in various pretreatments showed that the highest GE was noticed in T12 (6.85), followed by T11 (5.81) and

**Peak Value (PV)**: Peak value was highest in T12 treatment (12.50), and least

**Germination Value (GV)**: The highest germination value was also observed in

**Germination Percentage (GP)**: 100% germination was achieved in T12 treatment (acid scarification for 30 minutes) followed by 97% in T11 and T10 (acid scarification for 20 and 10 minutes). However, 10% was noticed in TI (no treatment) and T3 (soaked in water for 48 h) followed by 13.3% in T2 (soaked in water for 24 h), T8 (500 ppm GA3) and T9 (1000 ppm GA3). Hot water treatments followed by water soaking (T6 and T7) exhibited about 33% germination, which

*ANOVA table—level of significance on mean values of seed germination under different pretreatments.*

was better than mere hot water treatment (T4, 20%, and T5, 27%).

Within groups 2669.618 202 13.216

T11 (09 days) and T10 (11 days) and the highest in T2 (26 days).

T10 (4.48). The least GE is in T3 (0.29) followed by T1 (0.30).

value (0.56) was noticed in treatments T1, T3, T8 and T9.

T12 (208.36), followed by T11 (116.31) and T10 (58.63).

treatment T12 (16.67), and the lowest value was in T3 treatment (0.43).

value was in seeds without any treatment (T1).

*DOI: http://dx.doi.org/10.5772/intechopen.84970*

T1, T2, T5, T8 and T9 treatments.

**Table 3.**

**Figure 1.**

Total 7002.393 213

#### **Table 2.**

*Seed characteristics of B. malabarica.*

*Seed Characteristics and Germination Behaviour of* Bauhinia malabarica *Roxb. DOI: http://dx.doi.org/10.5772/intechopen.84970*

#### **Figure 1.**

*Horticultural Crops*

**3.2 Maturity index**

**3.3 Pod/seed characteristics**

**3.4 Seed germination**

value are given in **Table 4**.

*Pod/seed maturity indices of B. malabarica.*

and hardness (pods and seeds become hardest).

germination, etc. are presented in **Table 2**.

The optimum maturity indices for seed collection of *B. malabarica* were presented in **Table 1**. The maturity indices identified for determining optimum period for seed collection were colour of pods (yellowish-green turned to blackish-green), leaf number (minimum number of leaves), dehydration (pods become dehydrated)

Colour, shape, size, type, weight, number of seeds per pod and per kg, type of

**Figure 1** depicts germination pattern of seeds under various pretreatments. Seed

Germination-related parameters under different pretreatments such as germination initial time, germination percentage, germination duration, mean germination time, mean daily germination, germination energy, peak value and germination

germination among treatments was significantly different (**Table 3**).

Fruit Colour Yellowish-green turned to blackish-green Water content Very less Hardness Hardened

Hardness Hardened

**Character Variable Nature** Fruit Type Pod

**Character Variable Nature**

Seed Colour Dark brown

Leaf Number Minimum

Seed No. of seeds/fruit 10 ± 2

Germination Type Epigeal

Colour (young) Green

Colour (mature) Blackish brown Size (cm) 15.25 ± 4.75 × 2.15 ± 0.25

Wall type Dry, semihard

Colour Dark brown Shape Broad elliptic/oblong Size (mm) .02 ± 1.11 × 6.14 ± 0.88

Wall type Dry, hard Weight 7092 ± 50

**6**

**Table 2.**

**Table 1.**

*Seed characteristics of B. malabarica.*

*Seed germination pattern under different treatments. Note: T1, control; T2, soaked in water for 24 h; T3, soaked in water for 48 h; T4, soaked in hot water for 2 min; T5, soaked in hot water for 5 min; T6, soaked in hot water for 2 min + soaked in water for 24 h; T7, soaked in hot water for 5 min + soaked in water for 24 h; T8, soaked in gibberellic acid (GA3—500 ppm) for 2 h; T9, soaked in gibberellic acid (GA3—1000 ppm) for 2 h; T10, acid treatment (conc. H2SO4) for 10 min; T11, acid treatment (conc. H2SO4) for 20 min; T12, acid treatment (conc. H2SO4) for 30 min.*


#### **Table 3.**

*ANOVA table—level of significance on mean values of seed germination under different pretreatments.*

**Germination Initial Time (GIT)**: Germination initial time was 2 days after sowing in the treatments T4, T10, T11 and T12, whereas it was 5 days after sowing in T1, T2, T5, T8 and T9 treatments.

**Germination Percentage (GP)**: 100% germination was achieved in T12 treatment (acid scarification for 30 minutes) followed by 97% in T11 and T10 (acid scarification for 20 and 10 minutes). However, 10% was noticed in TI (no treatment) and T3 (soaked in water for 48 h) followed by 13.3% in T2 (soaked in water for 24 h), T8 (500 ppm GA3) and T9 (1000 ppm GA3). Hot water treatments followed by water soaking (T6 and T7) exhibited about 33% germination, which was better than mere hot water treatment (T4, 20%, and T5, 27%).

**Germination Duration (GD)**: Seed germination started 2 days after sowing and completed in 28 days. The least GD noticed in treatment T12 (06 days) followed by T11 (09 days) and T10 (11 days) and the highest in T2 (26 days).

**Mean Germination Time (MGT)**: Mean germination time was more in T12 treatment (21.67) where the treatment gave 100% germination, whereas the least value was in seeds without any treatment (T1).

**Mean Daily Germination (MDG)**: Mean daily germination was highest in the treatment T12 (16.67), and the lowest value was in T3 treatment (0.43).

**Germination Energy (GE)**: Germination energy in various pretreatments showed that the highest GE was noticed in T12 (6.85), followed by T11 (5.81) and T10 (4.48). The least GE is in T3 (0.29) followed by T1 (0.30).

**Peak Value (PV)**: Peak value was highest in T12 treatment (12.50), and least value (0.56) was noticed in treatments T1, T3, T8 and T9.

**Germination Value (GV)**: The highest germination value was also observed in T12 (208.36), followed by T11 (116.31) and T10 (58.63).


**Note:** *T1, control; T2, soaked in water for 24 h; T3, soaked in water for 48 h; T4 soaked in hot water for 2 min; T5 soaked in hot water for 5 min; T6, soaked in hot water for 2 min + soaked in water for 24 h; T7, soaked in hot water for 5 min + soaked in water for 24 h; T8, soaked in GA3 (500 ppm) for 2 h; T9, soaked in GA3 (1000 ppm) for 2 h; T10, acid treatment (conc. H2SO4) for 10 min; T11, acid treatment (conc. H2SO4) for 20 min; T12, acid treatment (conc. H2SO4) for 30 min; GIT, germination initial time; GP, germination percentage; GD, germination duration; MGT, mean germination time; MDG, mean daily germination; GE, germination energy; PV, peak value; GV, germination value.*

#### **Table 4.**

*Seed germination-related parameters under different treatments.*

#### **4. Discussion**

Maturity indices help to collect seeds with maximum viable seeds. The optimum maturity indices of *B. malabarica* identified in the present study were the colour of pod turned from yellowish-green to blackish-brown, minimum number of leaves, pods become dehydrated and the pods and seeds become hardest. A previous study reported that seed germination of *Albizia lebbeck* significantly influenced by date of pod collection [25]. Seed weight of *B. malabarica* in the present study showed 7092 ± 50 seeds per kilogram. However, in an earlier report, it is 1100–2600 seeds per kilogram [26]. Seed size is usually related with its vigour and a measure of potential performance; hence, seed weight is significant. Information regarding seed weight and moisture content is helpful for nursery practices and research-oriented studies and also an updating of the earlier information regarding seed weight and moisture content and seed characteristics.

The present study indicated that the highest germination was recorded in acid treatments (acid scarification for 30, 20 and 10 min). All other treatments exhibited poor performance in germination. Germination initiation period was minimum in the treatments T4, T10, T11 and T12, whereas maximum was in T1, T2, T5, T8 and T9 treatments. The lowest germination initial time shows speedy initiation of germination among pretreatments.

Estimation of germination percentage is the best tool to explain seed viability of a particular lot. Seeds treated with concentrated sulphuric acid for 30 minutes (T12) resulted in very high germination rate (100%) than that of other treatments. The study also revealed that the hormonal treatment had no significant role on seed germination of *B. malabarica*. A similar result was reported in an earlier study on *B. rufescens* after acid treatment [17]. Better performance in acid scarification on seed germination was reported in seeds of many species having hard seed coat [27–29]. Germination percent

**9**

**5. Conclusions**

*Seed Characteristics and Germination Behaviour of* Bauhinia malabarica *Roxb.*

(PV) and germination values were also highest in the treatment T12.

and GV show the enhanced germination and reduced duration.

high germination value indicates the germination power of seeds.

is useful for computing seed requisite for desired number of plants. Pretreatment with

Germination duration (GD) is helpful to understand the duration required for completing the process of germination. Germination duration in the present study varied with treatments (06–26 days). The study showed that the seeds scarified with concentrated sulphuric acid for 30 min helped to reduce germination period into 6 days compared to 26 days in seeds soaked in water for 24 h. The lowest GD shows the minimum period required to complete germination among pretreatments. Mean germination time (MGT) is the indicative of emergence performance of seed lots. Mean germination time (MGT) was highest in seeds treated with concentrated sulphuric acid for 30 min (21.67) where the treatment gave 100% germination, whereas the least value was in seeds without any treatment (T1). Similarly, mean daily germination (MDG), germination energy (GE), peak value

Mean germination time and mean daily germination are used as a gauge of the rate and time spread of germination. High MGT and MDG values indicate high germinability of seed lots due to pretreatments. Peak value indicates the maximum germination rate in a particular day, and germination value is the expected seedlings in the field or nursery. Germination energy and germination value are the easier way to understand the rate of germination and period of germination. Highest GE

The study resulted in scarification of seeds by concentrated sulphuric acid for 30 min which was the best pre-sowing treatments for enhancing seed germination and reduce germination period in *B. malabarica*. Previous studies showed that acid scarification is the best pretreatment to improve germination of seeds with hard seed coat [27–29]. Similarly, parameters like germination percentage, germination energy, mean germination time, mean daily germination, peak value and germination value were also highest in the acid treatment for 30 min compared to other treatments. High values of parameters indicate the germination potential of the seeds. Germination energy/germination value is the tool for indexing the speed and completeness of seed germination [22]. High germination energy and germination value show the effects of pretreatment on seed germination. Period of seed germination in *B. malabarica* is reported in an earlier study with 6–30 days and hot water treatment for 1 min followed by soaking in cold water for 24 hours as the best pretreatment [26]. Similarly, a high rate of germination observed in seeds of *Hippophae salicifolia* treated with thiourea [30]. The Forest Research Institute, India, reported only a low rate of seed germination (14–18%) in *B. malabarica* [31]. However, the present study revealed that the concentrated sulphuric acid treatment for 30 min shall reduce germination period (6 days) and increase germination rate (100%). An earlier study on *Macaranga peltata* revealed that combination of concentrated sulphuric acid and gibberellic acid resulted in improved germination rate [32].

The study gave update to seed characteristics and germination behaviour of *B. malabarica*. The investigation has documented maturity indices for determining optimum seed collection period of the species *B. malabarica*, and the data is very helpful for further seed biological studies and experiments on the species. The study suggested optimum period for collection of seed of *B. malabarica* is when yellowishgreen colour of pods turned to blackish-green, dehydrated and hard. The study recommended that concentrated sulphuric acid treatment for 30 min is the best pretreatment for enhancing seed germination and reducing germination period.

*DOI: http://dx.doi.org/10.5772/intechopen.84970*

#### *Seed Characteristics and Germination Behaviour of* Bauhinia malabarica *Roxb. DOI: http://dx.doi.org/10.5772/intechopen.84970*

is useful for computing seed requisite for desired number of plants. Pretreatment with high germination value indicates the germination power of seeds.

Germination duration (GD) is helpful to understand the duration required for completing the process of germination. Germination duration in the present study varied with treatments (06–26 days). The study showed that the seeds scarified with concentrated sulphuric acid for 30 min helped to reduce germination period into 6 days compared to 26 days in seeds soaked in water for 24 h. The lowest GD shows the minimum period required to complete germination among pretreatments. Mean germination time (MGT) is the indicative of emergence performance of seed lots. Mean germination time (MGT) was highest in seeds treated with concentrated sulphuric acid for 30 min (21.67) where the treatment gave 100% germination, whereas the least value was in seeds without any treatment (T1). Similarly, mean daily germination (MDG), germination energy (GE), peak value (PV) and germination values were also highest in the treatment T12.

Mean germination time and mean daily germination are used as a gauge of the rate and time spread of germination. High MGT and MDG values indicate high germinability of seed lots due to pretreatments. Peak value indicates the maximum germination rate in a particular day, and germination value is the expected seedlings in the field or nursery. Germination energy and germination value are the easier way to understand the rate of germination and period of germination. Highest GE and GV show the enhanced germination and reduced duration.

The study resulted in scarification of seeds by concentrated sulphuric acid for 30 min which was the best pre-sowing treatments for enhancing seed germination and reduce germination period in *B. malabarica*. Previous studies showed that acid scarification is the best pretreatment to improve germination of seeds with hard seed coat [27–29]. Similarly, parameters like germination percentage, germination energy, mean germination time, mean daily germination, peak value and germination value were also highest in the acid treatment for 30 min compared to other treatments. High values of parameters indicate the germination potential of the seeds. Germination energy/germination value is the tool for indexing the speed and completeness of seed germination [22]. High germination energy and germination value show the effects of pretreatment on seed germination. Period of seed germination in *B. malabarica* is reported in an earlier study with 6–30 days and hot water treatment for 1 min followed by soaking in cold water for 24 hours as the best pretreatment [26]. Similarly, a high rate of germination observed in seeds of *Hippophae salicifolia* treated with thiourea [30]. The Forest Research Institute, India, reported only a low rate of seed germination (14–18%) in *B. malabarica* [31]. However, the present study revealed that the concentrated sulphuric acid treatment for 30 min shall reduce germination period (6 days) and increase germination rate (100%). An earlier study on *Macaranga peltata* revealed that combination of concentrated sulphuric acid and gibberellic acid resulted in improved germination rate [32].

#### **5. Conclusions**

The study gave update to seed characteristics and germination behaviour of *B. malabarica*. The investigation has documented maturity indices for determining optimum seed collection period of the species *B. malabarica*, and the data is very helpful for further seed biological studies and experiments on the species. The study suggested optimum period for collection of seed of *B. malabarica* is when yellowishgreen colour of pods turned to blackish-green, dehydrated and hard. The study recommended that concentrated sulphuric acid treatment for 30 min is the best pretreatment for enhancing seed germination and reducing germination period.

*Horticultural Crops*

**4. Discussion**

**Table 4.**

and moisture content and seed characteristics.

*Seed germination-related parameters under different treatments.*

germination among pretreatments.

Maturity indices help to collect seeds with maximum viable seeds. The optimum maturity indices of *B. malabarica* identified in the present study were the colour of pod turned from yellowish-green to blackish-brown, minimum number of leaves, pods become dehydrated and the pods and seeds become hardest. A previous study reported that seed germination of *Albizia lebbeck* significantly influenced by date of pod collection [25]. Seed weight of *B. malabarica* in the present study showed 7092 ± 50 seeds per kilogram. However, in an earlier report, it is 1100–2600 seeds per kilogram [26]. Seed size is usually related with its vigour and a measure of potential performance; hence, seed weight is significant. Information regarding seed weight and moisture content is helpful for nursery practices and research-oriented studies and also an updating of the earlier information regarding seed weight

**Treatment GIT GP GD MGT MDG GE PV GV** T1 5 10 22 1.77 0.45 0.30 0.56 0.25 T2 5 13.3 26 2.23 0.51 0.42 1.11 0.57 T3 4 10 23 1.83 0.43 0.29 0.56 0.24 T4 2 20 25 3.28 0.80 0.59 1.11 0.89 T5 5 26.7 25 4.08 1.07 0.81 1.11 1.19 T6 4 33.3 25 6.56 1.33 0.78 0.67 0.89 T7 4 33.3 24 5.54 1.39 0.92 1.11 1.54 T8 5 13.3 21 2.57 0.63 0.37 0.56 0.35 T9 5 13.3 21 2.57 0.63 0.37 0.56 0.35 T10 2 96.7 11 18.27 8.79 4.48 6.67 58.63 T11 2 96.7 9 17.56 10.74 5.81 10.83 116.31 T12 2 100 6 21.67 16.67 6.85 12.50 208.36 **Note:** *T1, control; T2, soaked in water for 24 h; T3, soaked in water for 48 h; T4 soaked in hot water for 2 min; T5 soaked in hot water for 5 min; T6, soaked in hot water for 2 min + soaked in water for 24 h; T7, soaked in hot water for 5 min + soaked in water for 24 h; T8, soaked in GA3 (500 ppm) for 2 h; T9, soaked in GA3 (1000 ppm) for 2 h; T10, acid treatment (conc. H2SO4) for 10 min; T11, acid treatment (conc. H2SO4) for 20 min; T12, acid treatment (conc. H2SO4) for 30 min; GIT, germination initial time; GP, germination percentage; GD, germination duration; MGT, mean germination time; MDG, mean daily germination; GE, germination energy; PV, peak value; GV, germination value.*

The present study indicated that the highest germination was recorded in acid treatments (acid scarification for 30, 20 and 10 min). All other treatments exhibited poor performance in germination. Germination initiation period was minimum in the treatments T4, T10, T11 and T12, whereas maximum was in T1, T2, T5, T8 and T9 treatments. The lowest germination initial time shows speedy initiation of

Estimation of germination percentage is the best tool to explain seed viability of a particular lot. Seeds treated with concentrated sulphuric acid for 30 minutes (T12) resulted in very high germination rate (100%) than that of other treatments. The study also revealed that the hormonal treatment had no significant role on seed germination of *B. malabarica*. A similar result was reported in an earlier study on *B. rufescens* after acid treatment [17]. Better performance in acid scarification on seed germination was reported in seeds of many species having hard seed coat [27–29]. Germination percent

**8**

## **Acknowledgements**

We are grateful to the Kerala Forest Research Institute for scientific support. We thank Ms. Lakshmikutty VA and Mr. Suresh MK, the support staffs of Kerala Forest Seed Centre, for providing help and cooperation during the experiment.

## **Author details**

Pazhayaveetil Kuttanpillai Chandrasekhara Pillai\*, Sanal Chalil Viswanath, Thoduvayil Karunakaran Hrideek and Aviyan Hari Jiji Kerala Forest Research Institute, Thrissur, Kerala, India

\*Address all correspondence to: pkcpillai@gmail.com

© 2019 The Author(s). Licensee IntechOpen. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/ by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

**11**

*Seed Characteristics and Germination Behaviour of* Bauhinia malabarica *Roxb.*

[10] Darne P, Mehta M, Dubey P, Prabhune A. *Bauhinia* seed oil, a novel substrate for sophorolipid

and Pharmaceutical Sciences.

Medicines. 2008;**62**:364-365

[12] Kittakoop P, Kirtikara K, Tanticharoen M, Thebtaranonth Y. Antimalarial preracemosols A and B, possible biogenetic precursors of racemosol from *Bauhinia malabarica* Roxb. Phytochemistry.

2014;**3**(11):792-804

2000;**55**(4):349-352

2014;**4**(32):48-56

1993;**44**:291-298

1983;**85**(4):169-170

1987;**27**(1):139-153

[16] El-Sharkawi HM, Farghali KA. Interactive effects of nitrogen, water stress and temperature in the germination of *Bauhinia* seeds. Phyton.

[17] Asiedu JBK, Asare-Bediako E, Taah KJ, Buah JN. Effect of pre-sowing treatments on seed germination and

production. World Journal of Pharmacy

[11] Kaewamatawong R, Kitajima M, Kogure N, Takayama H. Flavanols from *Bauhinia malabarica*. Journal of Natural

[13] Thenmozhi K, Karthika K, Manian S, Paulsamy S. Studies on in vitro antioxidant potential of pod and seed parts of *Bauhinia malabarica* Roxb. Asian Journal of Biomedical and Pharmaceutical Sciences.

[14] Vijayakumari K, Sinddhuraju P, Janardhanan K. Chemical composition and nutrient potential of the tribal pulse (*Bauhinia malabarica* Roxb.). Plant Foods for Human Nutrition.

[15] Zaka S, Saleem M, Shakir N, Khan SA. Fatty acid composition of *Bauhinia variegata* and *Bauhinia malabarica* seed oils- comparison of their physiochemical properties. Wiley InterScience.

*DOI: http://dx.doi.org/10.5772/intechopen.84970*

[1] Roxburgh W. *Bauhinia*. In: Flora Indica. Calcutta, London: Carey; 1832.

[3] Albert S, Sharma B. Comparative foliar micromorphological studies of some *Bauhinia* (Leguminosae) species. Turkish Journal of Botany.

[4] Ponomarenko SF, Pavlova ME. Seed structure in the genus *Bauhinia* L. (fam. Fabaceae Lindlkm m.) in the context of taxonomy. Biology Bulletin of the Russian Academy of Sciences.

[5] Khan D, Zaki MJ. Size variation and surface structure of pods and seeds of *Bauhinia racemosa* Lamk. FUUAST Journal of Biology. 2016;**6**(1):73-80

[6] Anhwange BA, Ajibola VO, Oniye SJ. Nutritional potential of the seeds of *Bauhinia monandra* (Linn). Journal of Food Technology. 2005;**3**(2):204-208

[7] Jain S, Shrivastava BK. Structural determination of seed polysaccharide of *Bauhinia racemosa* by methylation studies. Oriental Journal of Chemistry.

[8] Sinha KN, Singh M, Kumar C. Electrophoretic study of seed storage protein in five species in Bauhinia. IOSR Journal of Pharmacy and Biological

[2] Bandyopadhyay S, Pathak MK. Neotypification of *Bauhinia malabarica* Roxb. (Fabaceae). Candollea. 2013;**68**(2):193-195

**References**

pp. 318-332

2013;**37**:276-281

2003;**30**(4):361-369

2005;**21**(3):601-602

Sciences. 2012;**4**(2):8-11

2011;**38**(2):242-251

[9] Phansri K, Sarthima R, Thammasirirak S, Boonchalee P, Khammuang S. Antibacterial activity of *Bauhinia acuminata* L. seed protein extract with low hemolytic activity against human erythrocytes. Chiang Mai Journal of Science.

*Seed Characteristics and Germination Behaviour of* Bauhinia malabarica *Roxb. DOI: http://dx.doi.org/10.5772/intechopen.84970*

#### **References**

*Horticultural Crops*

**Acknowledgements**

**10**

**Author details**

provided the original work is properly cited.

© 2019 The Author(s). Licensee IntechOpen. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/ by/3.0), which permits unrestricted use, distribution, and reproduction in any medium,

Pazhayaveetil Kuttanpillai Chandrasekhara Pillai\*, Sanal Chalil Viswanath,

We are grateful to the Kerala Forest Research Institute for scientific support. We thank Ms. Lakshmikutty VA and Mr. Suresh MK, the support staffs of Kerala Forest

Seed Centre, for providing help and cooperation during the experiment.

Thoduvayil Karunakaran Hrideek and Aviyan Hari Jiji Kerala Forest Research Institute, Thrissur, Kerala, India

\*Address all correspondence to: pkcpillai@gmail.com

[1] Roxburgh W. *Bauhinia*. In: Flora Indica. Calcutta, London: Carey; 1832. pp. 318-332

[2] Bandyopadhyay S, Pathak MK. Neotypification of *Bauhinia malabarica* Roxb. (Fabaceae). Candollea. 2013;**68**(2):193-195

[3] Albert S, Sharma B. Comparative foliar micromorphological studies of some *Bauhinia* (Leguminosae) species. Turkish Journal of Botany. 2013;**37**:276-281

[4] Ponomarenko SF, Pavlova ME. Seed structure in the genus *Bauhinia* L. (fam. Fabaceae Lindlkm m.) in the context of taxonomy. Biology Bulletin of the Russian Academy of Sciences. 2003;**30**(4):361-369

[5] Khan D, Zaki MJ. Size variation and surface structure of pods and seeds of *Bauhinia racemosa* Lamk. FUUAST Journal of Biology. 2016;**6**(1):73-80

[6] Anhwange BA, Ajibola VO, Oniye SJ. Nutritional potential of the seeds of *Bauhinia monandra* (Linn). Journal of Food Technology. 2005;**3**(2):204-208

[7] Jain S, Shrivastava BK. Structural determination of seed polysaccharide of *Bauhinia racemosa* by methylation studies. Oriental Journal of Chemistry. 2005;**21**(3):601-602

[8] Sinha KN, Singh M, Kumar C. Electrophoretic study of seed storage protein in five species in Bauhinia. IOSR Journal of Pharmacy and Biological Sciences. 2012;**4**(2):8-11

[9] Phansri K, Sarthima R, Thammasirirak S, Boonchalee P, Khammuang S. Antibacterial activity of *Bauhinia acuminata* L. seed protein extract with low hemolytic activity against human erythrocytes. Chiang Mai Journal of Science. 2011;**38**(2):242-251

[10] Darne P, Mehta M, Dubey P, Prabhune A. *Bauhinia* seed oil, a novel substrate for sophorolipid production. World Journal of Pharmacy and Pharmaceutical Sciences. 2014;**3**(11):792-804

[11] Kaewamatawong R, Kitajima M, Kogure N, Takayama H. Flavanols from *Bauhinia malabarica*. Journal of Natural Medicines. 2008;**62**:364-365

[12] Kittakoop P, Kirtikara K, Tanticharoen M, Thebtaranonth Y. Antimalarial preracemosols A and B, possible biogenetic precursors of racemosol from *Bauhinia malabarica* Roxb. Phytochemistry. 2000;**55**(4):349-352

[13] Thenmozhi K, Karthika K, Manian S, Paulsamy S. Studies on in vitro antioxidant potential of pod and seed parts of *Bauhinia malabarica* Roxb. Asian Journal of Biomedical and Pharmaceutical Sciences. 2014;**4**(32):48-56

[14] Vijayakumari K, Sinddhuraju P, Janardhanan K. Chemical composition and nutrient potential of the tribal pulse (*Bauhinia malabarica* Roxb.). Plant Foods for Human Nutrition. 1993;**44**:291-298

[15] Zaka S, Saleem M, Shakir N, Khan SA. Fatty acid composition of *Bauhinia variegata* and *Bauhinia malabarica* seed oils- comparison of their physiochemical properties. Wiley InterScience. 1983;**85**(4):169-170

[16] El-Sharkawi HM, Farghali KA. Interactive effects of nitrogen, water stress and temperature in the germination of *Bauhinia* seeds. Phyton. 1987;**27**(1):139-153

[17] Asiedu JBK, Asare-Bediako E, Taah KJ, Buah JN. Effect of pre-sowing treatments on seed germination and

establishment of *Bauhinia rufescens*. International Journal of Agricultural Science. 2011;**6**(7):584-592

[18] Mwase WF, Mvula T. Effect of seed size and pretreatment methods of *Bauhinia thonningii* Schum. on germination and seedling growth. African Journal of Biotechnology. 2011;**10**(13):5143-5148

[19] ISTA. In: Kruse M, editor. Handbook on Seed Sampling. 2nd ed. Bassersdorf, Switzerland: International Seed Testing Association; 2004

[20] Bewley JD. Seed germination and dormancy. Plant Cell. 1997;**9**:1055-1066

[21] Coolbear P. Pre-sowing treatments to improve seed performance. In: Proceedings of Seed Symposium: Seed Development and Germination. Tauranga: New Zealand Society of Plant Physiologists and Agronomy Society of New Zealand; 1991, August 1991. pp. 69-76

[22] Xu Y, Cai N, He B, Zhang R, Zhao R, Zhao W, et al. Germination and early seedling growth of *Pinus densata* mast. Provenances. Journal of Forest Research. 2016;**27**(2):283-294

[23] Czabator FJ. Germination value: An index combining speed and completeness of pine seed germination. Forest Science. 1962;**8**:386-395

[24] Djavanshir K, Pourbeik H. Germination value—A new formula. Silvae Genetica. 1976;**25**:79-83

[25] Kindt R, Muasya S, Kimotho J, Waruhiu. Tree seed supplier's dictionary: Sources of seeds and microsymbionts. In: International Centre for Research in Agro-Forestry (ICRAF). Nairobi: Kenya; 1997

[26] Malla S, Shukla S, Chakravarty S. Standardized pod collection time, pod length and pre-sowing treatment of

*Albizia lebbeck* at Terai zone of West Bengal, India. Journal of Forestry Research. 2012;**21**(3):338-342

[27] Usberti R, Martins L. Sulphuric acid scarification effects on *Brachiaria brizantha*, *B. humidicola* and *Panicum maximum* seed dormancy release. Revista Brasileira de Sementes. 2007;**29**(2):143-147

[28] Soliman AS, Abbas MS. Effects of sulfuric acid and hot water pretreatments on seed germination and seedlings growth of *Cassia fistula* L. American Eurasian Journal of Agriculture and Environmental Science. 2013;**13**(1):7-15

[29] Kher MM, Nataraj M. Effect of sulfuric acid treatment on breaking of seed dormancy and germination of Indian doum palm, *Hyphaene dichotoma*, a threatened and endemic palm. Environmental and Experimental Biology. 2015;**13**:99-101

[30] Airi S, Bhatt ID, Bhatt A, Rawal RS, Dhar U. Variations in seed germination of *Hippophae salicifolia* with different presoaking treatments. Journal of Forestry Research. 2009;**20**(1):27-30

[31] FRI. Troup's the Silviculture of Indian trees. Vol. IV. Delhi: The Controller of Publications; 1983. pp. 168-170

[32] Rodrigues CR, Rodrigues BF. Enhancement of seed germination in *Macaranga peltata* for use in tropical forest restoration. Journal of Forestry Research. 2014;**25**(4):897-901

**13**

**Chapter 2**

**Abstract**

were 25.78 and 384 μmol m<sup>−</sup><sup>2</sup>

**1. Introduction**

s<sup>−</sup><sup>1</sup>

light compensation point, light saturation point

tion management especially in light management of *Cymbidium*.

**Keywords:** *Cymbidium faberi*, photosynthetic characteristics, Qinling mountains,

*Cymbidium faberi* (Orchidaceae) is one of the several traditional and famous

Wild populations of *C. faberi* are mainly distributed in the southern mountainous area of China. The Qinling Mountains support the most northern population of wild *Cymbidium* species in China, where light serves as one of the most important factors affecting its natural distribution, growth, and development. The Qinling Mountains, located at 32°40′–34°35′N and 105°30′–110°05′E, run through the

orchid flowers in China. The Chinese have cultivated orchids for more than 2500 years. Most scientists currently recognize seven *Cymbidium* species in China: *C. sinensis* (Jackson ex Andr.) Willd., *C. ensifolium* (L.) Sw., *C. goeringii* (Rchb. f.), *C. faberi* Rolfe., *C. kanrau* Makino., *C. lianpan* Tang and F.T. Wang ex Y.S. Wu, and *C. longibracteatum* W.S. Wu & S.C. Chen [1]. To date, the British Royal Horticultural Society has registered 227 hybrids derived from Chinese orchids. Chinese orchids have been used as parents in the breeding of *C. faberi* because it is easy to grow, exhibits various flower colors and types, and gives off a sweet fragrance.

*Junyang Song and Ning Zhang*

The Photosynthetic Characteristics

Qinling Mountains of Central China

The large flowers of orchids make them popular as cultivated plants. Seven species of orchids in the genus *Cymbidium* (Orchidaceae) have been crossbred to create more than 220 hybrids that serve as popular cultivated ornamentals. The present study examined the daily variation in the patterns of the net photosynthetic rate and the photosynthetic response of wild *Cymbidium faberi* in the Qinling Mountains in northwestern China. The photosynthetic characteristics of this species were studied under natural conditions with a portable photosynthesis system. Double peaks were observed in the net photosynthetic rate with one around 09:00 and another around 17:00 in spring, as well as one around 11:00 and another around 15:00 in winter. Midday depression of photosynthesis was observed in wild *C. faberi* plants around 13:00 in both spring and winter. The net photosynthetic rate was strongly positively correlated with both stomatal conductance (*R* = 0.913) and the transpiration rate (*R* = 0.659) and weakly negatively correlated with the intercellular carbon dioxide concentration (*R* = −0.094). The results show that the light compensation point (LCP) and the light saturation point (LSP) of wild *C. faberi*

, respectively. The result provides reference for cultiva-

of Wild *Cymbidium faberi* in the

#### **Chapter 2**

*Horticultural Crops*

establishment of *Bauhinia rufescens*. International Journal of Agricultural *Albizia lebbeck* at Terai zone of West Bengal, India. Journal of Forestry Research. 2012;**21**(3):338-342

[27] Usberti R, Martins L. Sulphuric acid scarification effects on *Brachiaria brizantha*, *B. humidicola* and *Panicum maximum* seed dormancy release. Revista Brasileira de Sementes.

[28] Soliman AS, Abbas MS. Effects of sulfuric acid and hot water pretreatments on seed germination and seedlings growth of *Cassia fistula* L. American Eurasian Journal of

Agriculture and Environmental Science.

[30] Airi S, Bhatt ID, Bhatt A, Rawal RS, Dhar U. Variations in seed germination of *Hippophae salicifolia* with different presoaking treatments. Journal of Forestry Research. 2009;**20**(1):27-30

[31] FRI. Troup's the Silviculture of Indian trees. Vol. IV. Delhi: The Controller of Publications; 1983.

[32] Rodrigues CR, Rodrigues BF. Enhancement of seed germination in *Macaranga peltata* for use in tropical forest restoration. Journal of Forestry

Research. 2014;**25**(4):897-901

[29] Kher MM, Nataraj M. Effect of sulfuric acid treatment on breaking of seed dormancy and germination of Indian doum palm, *Hyphaene dichotoma*, a threatened and endemic palm. Environmental and Experimental

2007;**29**(2):143-147

2013;**13**(1):7-15

Biology. 2015;**13**:99-101

pp. 168-170

[18] Mwase WF, Mvula T. Effect of seed size and pretreatment methods of *Bauhinia thonningii* Schum. on germination and seedling growth. African Journal of Biotechnology.

[19] ISTA. In: Kruse M, editor. Handbook on Seed Sampling. 2nd ed. Bassersdorf, Switzerland: International Seed Testing

[20] Bewley JD. Seed germination and dormancy. Plant Cell. 1997;**9**:1055-1066

[21] Coolbear P. Pre-sowing treatments to improve seed performance. In: Proceedings of Seed Symposium: Seed Development and Germination. Tauranga: New Zealand Society of Plant Physiologists and Agronomy Society of New Zealand; 1991, August 1991.

[22] Xu Y, Cai N, He B, Zhang R, Zhao R, Zhao W, et al. Germination and early seedling growth of *Pinus densata* mast. Provenances. Journal of Forest Research.

[23] Czabator FJ. Germination value: An index combining speed and

Forest Science. 1962;**8**:386-395

[24] Djavanshir K, Pourbeik H. Germination value—A new formula. Silvae Genetica. 1976;**25**:79-83

[25] Kindt R, Muasya S, Kimotho J, Waruhiu. Tree seed supplier's dictionary: Sources of seeds and microsymbionts. In: International Centre for Research in Agro-Forestry (ICRAF). Nairobi: Kenya; 1997

[26] Malla S, Shukla S, Chakravarty S. Standardized pod collection time, pod length and pre-sowing treatment of

completeness of pine seed germination.

Science. 2011;**6**(7):584-592

2011;**10**(13):5143-5148

Association; 2004

pp. 69-76

2016;**27**(2):283-294

**12**
