A taxonomic study on pollen morphology characteristics, chromosome karyotype characteristics, floral and fruit pigment compositions of 13 species and hybrids of Lonicera L.

doi:10.21203/rs.3.rs-1657930/v1

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A taxonomic study on pollen morphology characteristics, chromosome karyotype characteristics, floral and fruit pigment compositions of 13 species and hybrids of Lonicera L.

https://doi.org/10.21203/rs.3.rs-1657930/v1

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In this paper, the pollen morphology characteristics, chromosome karyotype characteristics, floral and fruity pigment compositions of 13 species and hybrids of Lonicera L. were analyzed by means of scanning electron microscopy, root tip squash method, HPLC-MS and GC-MS, and then they were classified separately to discuss their genetic relationship and taxonomic relationship.

According to the morphological taxonomy, Lonicera L. distributed in northern are divided into two categories by whether the branchlets are hollow or not. In this experiment, Lonicera chrysanthaTurcz. ex Ledeb., Lonicera maackii (Rupr.) Maxim., Lonicera ruprechtiana Regel., Lonicera tatarica Linn. and Lonicera korolkowi ‘Zabelii’. belongsto the category of hollow branchlets; and Lonicera edulisTurcz. and Lonicera tatarinowiiMaxim. belongs to the category of branchlets with solid white pith. Through the analysis of pollen morphology characteristics, chromosome karyotype characteristics, floral and fruit pigment compositions, and using the above four indexes for clustering, Lonicera ruprechtiana Regel. and Lonicera tatarica Linn. can always be clustered together. Five hybrids such as Lonicera korolkow‘Zabelii’.×Lonicera maackii (Rupr.) Maxim., Lonicera korolkow‘Zabelii’.×Lonicera tatarica Linn.No.8 etal. can basically clustered with their parents, which is conform to the morphological taxonomy. These results indicate that the analysis of pollen morphology characteristics, chromosome karyotype characteristics, floral and fruit pigment compositions, has certain value and significance for the study of system classification and genetic relationship of Lonicera L., and the conclusions of this study partially support morphological taxonomy. Among the four clusters, the clustering result of pollen morphology characteristics was in the highest agreement with morphological taxonomy.

13 species and hybrids of Lonicera L.

Pollen morphology characteristics

Chromosome karyotype characteristics

Floral pigment compositions

Fruit pigment compositions

Classification relations

Lonicera L. is rich in resources and widely distributed in north temperate zone. There are about 200 species of Lonicera L. in the world, 98 species in China and 11 species in Heilongjiang Province(YanchengTang and Liangqian Li 1994; Zhengyi Wu et al. 2003).Lonicera L. have ornamental values such as flowers, fruits, leaf colors and tree postures and have ecological adaptability such as cold tolerance, shade tolerance, humidity tolerance, drought tolerance and salt and alkali tolerance. Most of them also have medicinal and healthy care functions. Therefore, in recent years, the introduction, application and cross breeding of new varieties of Lonicera L. have been widely carried out, and a large number of new varieties superior to their parents have been cultivated. Beijing took the lead in introducing varieties such as "mint" Lonicera, "golden flame" jinghong Lonicera, "beauty" jinghong Lonicera and "scarlet" brown Lonicera from abroad, and carried out cross breeding with local Lonicera. L. After extensive introduction and breeding, many new cultivars and varieties of Lonicera L. were produced, resulting in unclear genetic relationship of some varieties or uncertain origin of some varieties, So the genetic relationship and taxonomic relationship of these Lonicera L. varieties are confused, and the taxonomic relationship between these Lonicera L. varieties can not be clarified by traditional morphological taxonomy. At present, scientific methods are indeed needed to explore the classification of these new Lonicera L. varieties.

The research showed that pollen, chromosome, floral pigment and fruit pigment of a species are all genetic material of this species, which can be stably transmitted to offspring, reflecting the genetic information of species and also are the basis for classification of species(Attar et al. 2018; Ullah et al. 2018; Xiong et al. 2019). Talebi, SM et al. analyzed the pollen morphology of 19 species of Onobrychis Miller(Fabaceae), and considered that their pollen morphology had taxonomic significance for the division of subfamilies, subgenera and species(Talebi et al. 2020).Xin Wang et al. analyzed the karyotypes of 7 species of Rudbeckia, and considered that their karyotypes were of great significance for identifying the genetic relationship and evolution degree of Rudbeckia (Xin Wang et al. 2021). Yizeng Lu analyzed the anthocyanin components of Camellia, and the domestic and foreign Camellia varieties were divided(Yizeng Lu et al. 2007). Shufang Yan et al. made a cluster analysis of the anthocyanins in the fruits of 18 species of Malus hupehensis, and providing a basis for the identification of Malus hupehensis(Shufang Yan et al. 2016).

Due to extensive introduction and breeding, the classification relationship of Lonicera L. varieties is unclear, and the classification of cultivars and varieties only by means of morphological and anatomical features has obvious limitations. Literature review shows that there is no report on the application of pollen characteristics, chromosome karyotype characteristics, floral and fruit pigments components clustering to identify and classify Lonicera L.in the meanwhile. This paper intends to analyze the genetic relationship of 13 species and hybrids of Lonicera L. in Heilongjiang Province by cluster analysis of pollen morphological characteristics, chromosome karyotype characteristics, floral and fruit pigments components. The phylogenetic relationship was determined and compared with morphological taxonomy in order to provide scientific methods for the classification of species and varieties of Lonicera L.

2.1. Experimental materials

13 species and hybrids of Lonicera L. (8 species, 5 varieties) were selected as experimental materials, which were from Heilongjiang Forest Botanical Garden and Northeast Agricultural University campus. Their growth environment was basically the same. They were 4-year-old, and these plants with good growth potential and pure flower color and fruit color (Table 1).

Table 1

Information of experimental materials
NO	Latin name	Typical morphological characteristics of flowers and fruits
1	Lonicera maackii (Rupr.) Maxim.	white corolla at first and then yellow red berry
2	Lonicera ferdinandiFranch.	white corolla at first and then yellow red berry
3	Lonicera korolkowi ‘Zabelii’.	purple-red corolla red berry
4	Lonicera tatarica Linn.	white corolla at first and then pink red berry
5	Lonicera ruprechtiana Regel.	white corolla at first and then yellow orange berry at first and then red
6	Lonicera chrysantha Turcz. ex Ledeb.	yellow-white corolla red berry
7	Lonicera tatarinowii Maxim.	deep purple corolla red berry
8	Lonicera edulis Turcz.	yellow-white corolla deep blue berry covered with white powder
9	Lonicera korolkow ‘Zabelii’.× Lonicera maackii (Rupr.) Maxim.	white corolla at first and then yellow red berry
10	Lonicera korolkow‘Zabelii’.× Lonicera tatarica Linn. No.1	white corolla at first and then yellow red berry
11	Lonicera korolkow‘Zabelii’.× Lonicera tatarica Linn. No.2	rose-red corolla red berry
12	Lonicera korolkow‘Zabelii’.× Lonicera tatarica Linn. No.4	pink corolla red berry
13	Lonicera korolkow‘Zabelii’.× Lonicera tatarica Linn. No.8	rose-red corolla, white in center red berry
^{(Note: Due to the abortion of Lonicera tatarinowii Maxim. in the Heilongjiang Forest Botanical Garden, its fruit cannot be collected, and the fruit color composition and cluster analysis did not include this species.)}

2.2. Experimental method

Morphological characteristics of Lonicera L. pollen: sampling and fixation, dehydration and replacement, drying and coating, measurement and photographing according to scanning electron microscope method(Al-Hakimi et al. 2018; Bosen Xu and Jing Yang 2007). In a sunny day morning from May 20–30 in 2020, 30 mature anthers (without crack) from the flowers about to open in Lonicera L. were collected. Before the sample being coated, the anthers were gently broken with a dissecting needle to pick out the pollen(Borsch et al. 2018). Thirteen pollen morphological indexes were selected, including 8 quantitative indexes (polar axis length, equatorial axis length, pollen size, polar axis length/equatorial axis length, germinating pore length, germinating pore width, average protrusion height, protrusion density) and 5 qualitative indexes (pollen shape, equatorial plane view, polar plane view, germinating pore shape, exine ornamentation). When the indexes are quantified, the quantitative indexes take their values, and the qualitative indexes are assigned value to 1, 2,...n, according to their different forms. In the calculation, the values of each index are directly input(Xiaoxiao Wu et al. 2018). Among them, ES-2030 (Hitachi) freeze-drying instrument was used for freeze-drying samples, E-1010 (Hitachi) ion sputtering coating instrument was used for plating metal film on the surface of samples, and S-3400n(Hitachi, Japan) scanning electron microscope was used for observing, measuring and photographing pollen morphological indexes.

Chromosome karyotype characteristics of Lonicera L.: Refer to the root tip squashing method for sampling, fixation, dissociation, staining, measuring and photographing(Parrish et al. 2021; Wu et al. 2021). When the root length of Lonicera L. seeds was up to 0.5-1.0 cm, 20 root tips were taken and pressed into slices; the numbers of clear and scattered chromosomes in the cells was counted when more than 85% of the cells have a constant and consistent chromosome numbers in 20 cells in each species Lonicera L., then the photos were taken under the 100 times oil lens(Ferreira et al. 2020; Ruiyang Chen 2003; Yonggang Qiao et al. 2020). The chromosome length and arm ratio were calculated and chromosome type and karyotype classification were judged according to the average chromosome of 10–20 cells selected; when the indexes are quantified, the quantitative indexes take their values, and the qualitative indexes are assigned value to 1, 2 ,...n, according to their different forms. In the calculation, the values of each indexes are directly input. When nikon-e200 biological microscope was used to examine the chromosomes, chromosome types, average relative length, average arm ratio and karyotype types were judged according to LiMaoxue, Levan and Stebbins respectively(Lee et al. 2020; Sinela et al. 2017; Wicker 1991) .

Floral and fruit pigment compositions of Lonicera L.: According to HPLC-MS(Chai et al. 2021), 13 kinds of fresh flowers and mature fruits of Lonicera L. were collected in a sunny morning from April to September in 2020.5g petal and 5g fruits were grounded into powder by liquid nitrogen in the mortars, then 0.1% acidic methanol solution (methanol: water = 80:20, v/v) was added. The filtrate was filtered by filter paper and 0.22µm microporous membrane and the components will be identified by machine after ultrasonic-assisted extraction at 40℃ for 30 min and extraction at room temperature in the dark conditions for 24 h.

The model of chromatographic column is ACQUITY UPLC BEH C₁₈ whose size is 100mm ×2.1mm×1.7µm. The detection was done for 25 minutes under column temperature 40℃, injection volume 5µL and flow rate 0.4 mL/min by mobile phase A(0.1% formic acid aqueous solution) and mobile phase B ( pure acetonitrile); The elution procedure is as follows:5%-15% concentration of B at 0–3 min; 15% concentration of B at 3-3.5 min;15%-30%concentration of B at 3.5-6 min;30% concentration of B at 6-6.5 min; 30%-70% concentration of B at 6.5–12 min; 70% concentration of B at 12-12.5 min; 70%-100% concentration of B at 12.5–18 min; 100% concentration of B at 18–25 min; 100%-5% concentration of B at 25-25.5min; 5% concentration of B at 25.5–29 min.

Mass spectrometry conditions of pigments: positive ion mode, scanning range of primary mass spectrometry and secondary mass spectrometry are 100–1200 M/Z and 50-1200 M/Z respectively, nitrogen is used in each gas path. The Scan Type of ESI-MS and ESI-MS² is TOF. The Ion Spray Voltage Floating(ISVF)of ESI-MS and ESI-MS² is 5500 V. The Source Gas 1༈GS1༉of ESI-MS and ESI-MS² is 50 psi. The Source Gas 2༈GS2༉of ESI-MS and ESI-MS² is 50 psi. The Curtain Gas༈CUR༉of ESI-MS and ESI-MS² is 35 psi. The Temperature༈TEM༉of ESI-MS and ESI-MS² is 550℃. The Declustering Potential༈DP༉of ESI-MS and ESI-MS² is 90 V. The Collision Energy༈CE༉of ESI-MS is 10V, and ESI-MS² is 35V.The Collision Energy Spread༈CES༉of ESI-MS² is 15V.

The results were analyzed by Masterview and Peakview software.

The assignment value method of non-quantitative character is adopted while quantifing pigment components, (0 = Absent, 1 = Present)

3.1. Taxonomic relationship of 13 species and hybrids of Lonicera L. based on pollen characteristics.

3.1.1. Pollen morphological characteristics and value assignments of 13 species and hybrids of Lonicera L.

The qualitative and quantitative morphological indexes of Lonicera L. pollen are shown in Table 2,3.

Table 2

Pollen morphological characteristics and value assignments of 13 species and hybrids of *Lonicera* L.
Latin name	Pollen shape	Equator view	Polar view	Germinating pore shape	Exine ornamentation
Lonicera maackii (Rupr.) Maxim.	near spherical 1	triangular circle 1	near circle 1	oblong and convexity1	unsmooth pollen exine surface, with uneven-distributed spinous ornamentation1
Lonicera ferdinandiFranch.	near spherical 1	triangular circle 1	near circle 1	oblong and convexity1	unsmooth pollen exine surface, with uneven-distributed spinous ornamentation1
Lonicera korolkowi ‘Zabelii’.	near spherical 1	triangular circle 1	near circle 1	oblong and convexity1	unsmooth pollen exine surface, with uneven-distributed spinous ornamentation1
Lonicera tatarica Linn.	near spherical 1	triangular circle 1	near circle 1	oblong and convexity1	unsmooth pollen exine surface, with uneven-distributed spinous ornamentation1
Lonicera ruprechtiana Regel.	near spherical 1	triangular circle 1	near circle 1	oblong and convexity1	unsmooth pollen exine surface, with uneven-distributed spinous ornamentation1
Lonicera chrysanthaTurcz. ex Ledeb.	near spherical 1	triangular circle 1	near circle 1	oblong and convexity1	unsmooth pollen exine surface, with uneven-distributed spinous ornamentation1
Lonicera tatarinowiiMaxim.	near spherical 1	triangular circle 1	near circle 1	oblong and convexity1	unsmooth pollen exine surface, with uneven-distributed spinous ornamentation1
Lonicera edulisTurcz.	near spherical 1	triangular circle 1	ellipse2	oblong and convexity1	unsmooth pollen exine surface, with uneven-distributed spinous ornamentation1
Lonicera korolkow‘Zabelii’.× Lonicera maackii (Rupr.) Maxim.	near spherical 1	triangular circle 1	near circle 1	oblong and convexity1	unsmooth pollen exine surface, with uneven-distributed spinous ornamentation1
Lonicera korolkow‘Zabelii’.× Lonicera tatarica Linn.No.1	near spherical 1	triangular circle 1	near circle 1	oblong and convexity1	unsmooth pollen exine surface, with uneven-distributed spinous ornamentation1
Lonicera korolkow‘Zabelii’.× Lonicera tatarica Linn. No.2	near spherical 1	triangular circle 1	near circle 1	oblong and convexity1	unsmooth pollen exine surface, with uneven-distributed spinous ornamentation1
Lonicera korolkow‘Zabelii’.× Lonicera tatarica Linn. No.4	near spherical 1	triangular circle 1	near circle 1	oblong and convexity1	unsmooth pollen exine surface, with uneven-distributed spinous ornamentation1
Lonicera korolkow‘Zabelii’.× Lonicera tatarica Linn. No.8	near spherical 1	triangular circle 1	near circle 1	oblong and convexity1	unsmooth pollen exine surface, with uneven-distributed spinous ornamentation1

Table 3

Pollen morphological characteristics of 13 species and hybrids of *Lonicera* L. (X¯±SD)1)
Latin name	Polar axis length P(µm)	equatorial axis lengthE(µm)	pollen sixze (P×E)(µm)	Polaraxis length/equatoria axis length (P/E)	germinating pore length(µm)	germinating pore width (µm)	average protrusion height(µm)	protrusiondensity (one/per area)
Lonicera maackii (Rupr.) Maxim.	46.80 ± 1.42	42.50 ± 2.33	46.80(± 1.42)×42.50(± 2.33)	1.10 ± 0.03	16.5 ± 0.41	3.55 ± 0.26	1.90 ± 0.16	19.50 ± 0.36
Lonicera ferdinandiFranch.	58.60 ± 2.77	53.50 ± 3.82	58.60(± 2.77)×53.50(± 3.82)	1.10 ± 0.03	18.8 ± 0.37	5.16 ± 0.16	1.50 ± 0.24	25.46 ± 0.33
Lonicera korolkowi ‘Zabelii’.	39.30 ± 3.83	37.43 ± 2.53	39.30(± 3.83)×37.43(2.53)	1.05 ± 0.04	12.3 ± 0.25	2.23 ± 0.10	2.00 ± 0.29	28.92 ± 0.39
Lonicera tatarica Linn.	41.20 ± 4.81	39.28 ± 3.54	41.23(± 4.81)×39.28(± 3.54)	1.05 ± 0.03	12.8 ± 0.30	2.57 ± 0.20	2.20 ± 0.29	29.00 ± 0.23
Lonicera ruprechtiana Regel.	42.10 ± 3.34	39.07 ± 1.99	42.17(± 3.34)×39.07(± 1.99)	1.08 ± 0.07	17.9 ± 0.31	1.95 ± 0.07	2.30 ± 0.22	24.56 ± 0.15
Lonicera chrysanthaTurcz. ex Ledeb.	40.30 ± 5.53	38.50 ± 2.04	40.30(± 5.53)×38.50(± 2.04)	1.05 ± 0.10	11.3 ± 0.23	3.20 ± 0.11	2.10 ± 0.22	23.04 ± 0.10
Lonicera tatarinowiiMaxim.	33.50 ± 3.60	31.80 ± 2.78	33.50(± 3.60)×31.80(± 2.78)	1.05 ± 0.02	0	0	1.10 ± 0.22	35.28 ± 0.19
Lonicera edulisTurcz.	40.90 ± 4.79	32.40 ± 2.96	40.90(± 2.78)×32.40(± 2.96)	1.26 ± 0.06	10.6 ± 0.27	1.89 ± 0.16	1.20 ± 0.22	29.51 ± 0.26
Lonicera korolkow‘Zabelii’.× Lonicera maackii (Rupr.) Maxim.	38.03 ± 3.91	36.13 ± 3.17	38.03(± 3.91)×36.13(± 3.17)	1.05 ± 0.02	13.6 ± 0.20	2.22 ± 0.04	2.60 ± 0.05	21.60 ± 1.93
Lonicera korolkow‘Zabelii’.× Lonicera tatarica Linn.No.1	38.90 ± 3.14	34.23 ± 1.76	38.90(± 3.14)×34.23(± 1.76)	1.14 ± 0.04	11.8 ± 0.41	2.21 ± 0.13	1.90 ± 0.16	31.00 ± 0.18
Lonicera korolkow‘Zabelii’.× Lonicera tatarica Linn. No.2	39.60 ± 2.41	35.23 ± 2.56	39.60(± 2.41)×35.23(± 2.56)	1.12 ± 0.02	14.3 ± 0.48	2.22 ± 0.11	2.50 ± 0.22	37.50 ± 0.26
Lonicera korolkow‘Zabelii’.× Lonicera tatarica Linn. No.4	36.43 ± 2.98	34.13 ± 2.44	36.43(± 2.98)×34.13(± 2.44)	1.07 ± 0.01	13.5 ± 0.30	2.06 ± 0.06	2.20 ± 0.14	40.52 ± 0.23
Lonicera korolkow‘Zabelii’.× Lonicera tatarica Linn. No.8	38.43 ± 4.05	38.03 ± 1.98	38.43(± 4.05)×38.03(± 1.98)	1.01 ± 0.06	13.4 ± 0.34	2.47 ± 0.10	2.20 ± 0.24	37.91 ± 0.11

The data in the table show that the pollens of 13 species(varieties) of Lonicera L. are near spherical, triangular circle in polar plane, and subround or ellipse in equatorial plane, with three-pored, furrow-shaped, ellipse and convex germination pore. Pollen exine surface is unsmooth, with uneven-distributed spinous ornamentation. The pollen is a medium type of pollen, with a size of 33.50×31.80 µm to 58.67×53.50 µm and P/E between 1.01 and 1.26. The length of polar axis is between 33.50 µm to 58.67 µm, the length of equatorial axis is between 31.80 µm to 53.50 µm, the size of germinating pore is 0 ~ 18.85 µm×0 ~ 5.16 µm. The pollen of Lonicera L. belongs to the more evolved type because its exine surface is not smooth, with uneven-distributed spinous ornamentation.

3.1.2. Cluster analysis of pollen morphological characteristics of 13 species and hybrids of Lonicera L.

13 pollen indexes of 13 species and hybrids of Lonicera L. were selected and the results of principal component analysis were carried out by non-quantitative trait assignment method (Table 4).

Table 4

Five principal components and its accumulative contribution rates
Principal components	Y1	Y2	Y3	Y4	Y5
Characteristic value Contribution rate (%)	2.79 34.872	1.726 21.581	1.089 13.607	0.95 11.871	0.755 9.436
Cumulative contribution rate (%)	34.872	56.453	70.061	81.932	91.368

As shown in Table 4, the cumulative contribution rate of principal components reached 91.37% through principal component analysis, which could represent most information of pollen traits of Lonicera L. and could be used as the inclusion items in cluster analysis.

Cluster analysis was conducted on the pollen morphological characteristics of 13 species and hybrids of Lonicera L. and taxonomic tree diagram was drawn (Fig. 1).

X-axis indicate genetic distance, Y-axis indicate 13 species and hybrids of Lonicera L.

Figure 1. The clustering map of Pollen characters of 13 species and hybrids of Lonicera L.

Clustering results show that at the genetic distance of 25.0, the 13 species are divided into four groups: the first group consists of Lonicera tatarica Linn., Lonicera chrysantha Turcz. ex Ledeb., Lonicera korolkowi ‘Zabelii’., Lonicera korolkow‘Zabelii’.×Lonicera tatarica Linn.No.8, Lonicera ruprechtiana Regel. and Lonicera korolkow‘Zabelii’.×Lonicera maackii (Rupr.) Maxim. The second group consists of Lonicera korolkow‘Zabelii’.×Lonicera tatarica Linn. No.2, Lonicera korolkow‘Zabelii’.×Lonicera tatarica Linn. No.4 and Lonicera korolkow‘Zabelii’.×Lonicera tatarica Linn. No.1. The third group only consists of Lonicera tatarinowiiMaxim..The fourth group contains Lonicera maackii (Rupr.) Maxim., Lonicera ferdinandiFranch., and Lonicera edulisTurcz..

The pollen of Lonicera L. is similar in shape and exine ornamentation, so it can be distinguished according to the quantitative indexes of pollen. In the first group, the polar axis length of Lonicera L. pollen all ranged from 38.03 µm to 42.17 µm, and P/E ranged from 1.01 µm to 1.08 µm. The pollen sizes of the second group are similar, P/E ranged from 1.07 µm to 1.14 µm. The average size (P × E) of pollen in the third group is smallest. In the fourth group, the average protrusion height of the pollen exine ornamentation was similar, which was less than 2.00µm.

3.2. Taxonomic relationship of 13 species and hybrids of Lonicera L. based on chromosome karyotype characteristics.

3.2.1. Chromosome karyotype characteristics and value assignments of 13 species and hybrids of Lonicera L.

The statistical results in Table 5. are as follows:

Table 5

The chromosome karyotype characteristics and value assignments of 13 species and hybrids of *Lonicera* L.
Latin Name	Karyotype formula	Average relative length /%	Average arm ratio	Asymmetrical karyotype coefficient/%	Arm ratio > 2 ratio/ %	Lt/St	Centromeric index	Karyotype
Lonicera maackii (Rupr.) Maxim.	12m + 6sm1	2.66–3.89	1.49	59.19	11.11	1.95	40.91	2A 1
Lonicera ferdinandiFranch.	10m + 8sm2	3.45–7.30	1.65	59.85	0.00	2.12	37.39	1B 2
Lonicera korolkowi ‘Zabelii’.	14m + 4sm3	2.41–8.05	1.50	59.82	11.11	3.34	40.79	2B 3
Lonicera tatarica Linn.	18m4	3.45–7.30	1.27	55.47	0.00	2.01	44.64	1B 2
Lonicera ruprechtiana Regel.	16m + 2sm5	2.78–5.76	1.48	56.43	0.00	2.07	40.58	1B 2
Lonicera chrysanthaTurcz. ex Ledeb.	8m + 10sm6	2.14–4.15	1.72	63.03	11.11	1.94	37.09	2A 1
Lonicera tatarinowiiMaxim.	14m + 4sm3	2.95–7.33	1.54	60.98	11.11	2.48	40.06	2B 3
Lonicera edulisTurcz.	14m + 4sm3	1.81–3.90	1.49	59.15	22.22	2.15	41.00	2B 3
Lonicera korolkow‘Zabelii’.× Lonicera maackii (Rupr.) Maxim.	14m + 4sm3	2.46–4.93	1.47	58.61	11.11	2.00	41.24	2B 3
Lonicera korolkow‘Zabelii’.× Lonicera tatarica Linn. No.1	14m + 4sm3	2.52–5.31	1.46	59.68	11.11	2.68	41.39	2B 3
Lonicera korolkow‘Zabelii’.× Lonicera tatarica Linn. No.2	16m + 2sm5	3.45–7.30	1.32	56.54	0.00	2.57	43.36	1B 2
Lonicera korolkow‘Zabelii’.× Lonicera tatarica Linn. No.4	14m + 4sm3	2.72–5.87	1.39	58.20	11.11	2.16	42.73	2B 3
Lonicera korolkow‘Zabelii’.× Lonicera tatarica Linn. No.8	14m + 4sm3	2.23–4.48	1.43	58.94	0.00	2.01	41.64	1B 2

The chromosome number of 13 species and hybrids of Lonicera L. in the experiment is 2n = 18, which is composed of middle centromeric chromosome (m) and near middle centromeric chromosome (sm). Because of primitive 1B, 2B and 2A of karyotype types, indicating that 13 species and hybrids of Lonicera L. is not evolution. There are on no obvious evolutionary difference between 13 species.

3.2.2. Cluster analysis of chromosome karyotype characteristics of 13 species and hybrids of Lonicera L.

Three principal components were analysised according to the assignment volue method of non-quantitative traits based on nine chromosomes indexes of 13 species and hybrids of Lonicera L. (Table 6).

Table 6

Three principal components and its accumulative contribution rates
The principal component	Y1	Y2	Y3
Characteristic value Contribution rate (%)	3.235 53.916	1.426 23.76	1.013 16.882
Cumulative contribution rate(%)	53.916	77.678	94.561

As shown in Table 6., the contribution rate of principal components is 94.56%, indicating that the information contain in principal components could fully reflect most of the information of the karyotype parameters of Lonicera L., and could be used as the inclusion items in cluster analysis.

Cluster analysis was conducted on the chromosome karyotype characteristics of 13 species and hybrids of Lonicera L. and taxonomic tree diagram was drawn (Fig. 2).

Clustering results show that at the genetic distance of 25.0, the 13 species are divided into three groups: the first group consists of Lonicera korolkowi ‘Zabelii’., Lonicera korolkow‘Zabelii’.×Lonicera tatarica Linn.No.1, Lonicera maackii (Rupr.) Maxim., Lonicera korolkow‘Zabelii’.×Lonicera maackii (Rupr.) Maxim., Lonicera tatarinowiiMaxim., Lonicera korolkow‘Zabelii’.×Lonicera tatarica Linn.No.4 and Lonicera chrysanthaTurcz. ex Ledeb.. The second group only consists of Lonicera edulisTurcz.. The third group consists of Lonicera tatarica Linn., Lonicera korolkow‘Zabelii’.×Lonicera tatarica Linn.No.2, Lonicera korolkow‘Zabelii’.×Lonicera tatarica Linn.No.8, Lonicera ruprechtiana Regel. and Lonicera ferdinandiFranch..

The average arm ratio of Lonicera L. in the first group is relatively large, and the ratio of arm ratio > 2 is 11.11. The karyotype of Lonicera L. in the third group are 1B, and the ratio of arm ratio > 2 is 0.

3.3. Taxonomic relationship of 13 species and hybrids of Lonicera L. based on floral pigment compositions.

3.3.1. Floral pigment compositions analysis and value assignments of 13 species and hybrids of Lonicera L.

26 floral pigment compositions were identified from 13 species and hybrids of Lonicera L. and non-quantitative trait assignment method was adopted (including a certain composition being assigned as 1, excluding a certain composition being assigned as 0). The results are shown in Table 7.

Table 7

The value assignments of floral pigment compositions 13 species and hybrids of *Lonicera* L. (0 = Absent, 1 = Present)
Lonicera L. species	1	2	3	4	5	6	7	8	9	10	11	12	13	14	15	16	17	18	19	20	21	22	23	24	25	26
Lonicera maackii (Rupr.) Maxim.	1	1	1	1	1	1	1	1	1	0	0	0	0	0	0	1	0	0	0	0	0	0	0	0	0	0
Lonicera ferdinandiFranch.	1	0	1	1	1	0	1	1	1	1	1	1	0	1	1	0	0	0	1	0	1	0	0	0	0	0
Lonicera korolkowi ‘Zabelii’.	1	0	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	0	0	0	0	0	0	0
Lonicera tatarica Linn.	1	0	0	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	0	0	0	0	0	0
Lonicera ruprechtiana Regel.	1	0	1	1	1	1	1	1	1	1	1	1	1	1	0	1	1	1	0	1	0	0	0	0	0	0
Lonicera chrysanthaTurcz. ex Ledeb.	1	0	1	1	1	1	1	0	1	0	1	1	0	0	1	1	0	0	0	1	0	0	0	0	0	0
Lonicera tatarinowiiMaxim.	0	0	1	1	1	1	1	0	1	0	1	0	0	1	0	0	0	1	0	1	1	1	1	1	0	0
Lonicera edulisTurcz.	1	1	1	1	1	1	0	0	1	0	0	1	1	1	0	1	0	0	1	1	0	0	0	0	1	0
Lonicera korolkow‘Zabelii’.× Lonicera maackii (Rupr.) Maxim.	1	1	1	1	1	1	1	0	1	1	1	0	0	1	0	1	1	1	0	1	0	0	0	0	0	0
Lonicera korolkow‘Zabelii’.× Lonicera tatarica Linn.No.1	1	0	1	1	1	1	1	1	1	1	1	0	0	1	0	1	1	1	0	1	0	0	0	0	0	0
Lonicera korolkow‘Zabelii’.× Lonicera tatarica Linn.No.2	1	1	1	1	1	0	1	1	1	1	1	0	0	1	0	1	1	1	0	1	0	0	0	0	0	0
Lonicera korolkow‘Zabelii’.× Lonicera tatarica Linn.No.4	1	0	1	1	1	0	1	1	0	1	1	1	0	0	1	1	1	1	0	1	0	0	0	0	0	0
Lonicera korolkow‘Zabelii’.× Lonicera tatarica Linn.No.8	1	0	1	1	0	1	1	1	1	1	1	1	1	1	1	1	1	1	0	1	0	0	0	0	0	1

^{(Note: 1 quercetin−3−o−rutoside 2 hexyl quercetin−3−o−glucoside 3 quercetin−3−o−glucoside 4 kaempferol−3−o−rutoside 5 kaempferol−3−o−glucoside 6 geraniol−3−o−rutoside 7 Geranium rutoside 8 Geranium glucoside 9 paeoniflorin glucoside 10 vinblastin 11 isorhamnetin−7−o−rutoside 12 delphinidin Sophorae 13 quercetin−3−o−sambuoside 14 quercetin−3−o−diglucoside 15 3−hydroxyphlorin 16 Geranium −Malonyl glucoside 17 delphinidin − arabinose 18 cyanidin − Sophora 19 Petunia − Sophora 20 isorhamnetin−3−o−glucoside 21 isorhamnetin 22 kaempferol−3−o−sambuoside 23 ferulic acid glucoside 24 delphinidin − hexyl glucose 25 delphinidin − malonyl glucose 26 mallow glucoside)}

According to the results analysis in Table 7., 26 pigment components were detected from 13 species and hybrids of Lonicera L., including 16 flavonoid pigments and 10 anthocyanin pigments. Quercetin-3-o-rutoside, Quercetin-3-o-glucoside, Kaempferol-3-o-rutoside, Kaempferol-3-o-glucoside, Geranium-rutoside, Paeoniflorin-glucoside are common components in most Lonicera L. Paeoniflorin-glucoside is the main anthocyanin component in Lonicera L., which accounts for the largest proportion of anthocyanin in Lonicera L.as well as there generally contains flavonoid pigment components in flowers in Lonicera L.

3.3.2. Cluster analysis of floral pigment components of 13 species and hybrids of Lonicera L.

The floral pigments components of 13 species and hybrids of Lonicera L. were analyzed by principal component analysis (Table 8.).

Table 8

Eight principal components and its contribution rates
The principal component	Y1	Y2	Y3	Y4	Y5	Y6	Y7	Y8
Characteristic value contribution rate (%)	6.676	4.669	3.229	2.349	1.665	1.591	1.25	0.627
Contribution rate (%)	29.025	20.301	14.039	10.41	7.241	6.916	5.433	2.728
Cumulative contribution rate(%)	29.025	49.326	63.365	73.775	81.016	87.932	93.365	96.093

Table 8. shows that the contribution rate of principal components reaches 96.09%, which indicates that the information contained in principal components can fully reflect most of the information of floral pigment components of Lonicera L. and can be used as an entry option for cluster analysis.

Cluster analysis was conducted on the floral pigment components of 13 species and hybrids of Lonicera L. and taxonomic tree diagram was drawn (Fig. 3.).

Clustering results show that at the genetic distance of 10.0, the 13 species are divided into three groups: the first group consists of Lonicera ferdinandiFranch., Lonicera korolkow‘Zabelii’.×Lonicera tatarica Linn.No.4, Lonicera korolkow‘Zabelii’.×Lonicera tatarica Linn.No.1, Lonicera maackii (Rupr.) Maxim. and Lonicera chrysanthaTurcz. ex Ledeb. The second group consists of Lonicera korolkowi ‘Zabelii’., Lonicera ruprechtiana Regel., Lonicera korolkow‘Zabelii’.×Lonicera tatarica Linn.No.8, Lonicera korolkow‘Zabelii’.×Lonicera maackii (Rupr.) Maxim., Lonicera korolkow‘Zabelii’.×Lonicera tatarica Linn.No.2, Lonicera tatarica Linn. and Lonicera edulisTurcz.. The third group only consists of Lonicera tatarinowiiMaxim..

The first group contains quercetin-3-o-rutoside, kaempferol-3-o-rutoside, kaempferol-3-o-glucoside and geranium rutoside. The flower color of the second group is reddish and contains paeoniflorin glucoside, vinblastin, isorhamnetin-7-o-rutoside, geranium malonyl glucose and delphinidin arabinose, which contain more anthocyanin pigments than the other two groups. In the third group, Lonicera tatarinowiiMaxim. has different flower color from other Lonicera L. there are four anthocyanins in this group: orhamnetin, kaempferol-3-o-sambuoside, ferulic acid glucoside and delphinidin hexyl glucose.

3.4. Taxonomic relationship of 12 species and hybrids of Lonicera L. based on fruit pigment components.

3.4.1. Fruit pigment components analysis and value assignments of 12 species and hybrids of Lonicera L.

23 fruit pigment components were identified from 12 species and hybrids of Lonicera L. and non-quantitative trait assignment volue method was adopted (including a certain composition being assigned as 1, excluding a certain composition being assigned as 0). The results are shown in Table 9.

Table 9

The value assignments of fruit pigment compositions 12 species and hybrids of *Lonicera* L. (0 = Absent, 1 = Present)
Lonicera L. species	1	2	3	4	5	6	7	8	9	10	11	12	13	14	15	16	17	18	19	20	21	22	23
Lonicera maackii (Rupr.) Maxim.	1	1	1	1	1	1	1	1	1	1	1	1	0	0	0	0	0	0	0	0	0	0	0
Lonicera ferdinandiFranch.	1	1	0	1	0	1	0	0	0	0	1	1	1	0	0	0	0	0	0	0	0	0	0
Lonicera korolkowi ‘Zabelii’.	1	1	1	0	1	1	1	1	1	1	0	1	0	1	0	0	0	0	0	0	0	0	0
Lonicera tatarica Linn.	1	1	0	1	0	1	1	1	1	0	1	1	0	0	1	1	0	0	0	0	0	0	0
Lonicera ruprechtiana Regel.	1	1	1	1	1	1	1	1	0	0	1	1	0	1	0	1	0	0	0	0	0	0	0
Lonicera chrysanthaTurcz. ex Ledeb.	0	0	0	1	0	0	0	0	1	0	0	1	0	0	0	1	0	0	0	0	0	0	0
Lonicera edulisTurcz.	1	1	1	1	1	1	1	1	1	1	1	1	0	1	1	0	0	0	0	0	0	0	0
Lonicera korolkow‘Zabelii’.× Lonicera maackii (Rupr.) Maxim.	1	1	0	1	1	1	0	1	1	0	0	1	0	0	1	0	1	1	0	0	0	0	0
Lonicera korolkow‘Zabelii’.× Lonicera tatarica Linn.No.1	1	1	1	1	1	1	0	0	0	0	1	1	0	0	1	1	0	1	1	1	1	1	1
Lonicera korolkow‘Zabelii’.× Lonicera tatarica Linn.No.2	1	1	1	1	1	1	0	1	1	1	1	1	0	0	0	0	0	0	0	0	0	0	0
Lonicera korolkow‘Zabelii’.× Lonicera tatarica Linn.No.4	1	1	0	1	1	1	1	1	1	1	0	1	0	0	0	1	0	1	0	0	0	0	0
Lonicera korolkow‘Zabelii’.× Lonicera tatarica Linn.No.8	1	1	1	1	1	1	1	1	1	0	1	1	0	0	1	0	0	0	0	0	0	0	0
^{(Note: 1 vinblastin 2 quercetin−3−o−rutoside 3 hexyl quercetin−3−o−glucoside) 4 quercetin−3−o−glucoside 5 kaempferol−3−o−rutoside 6 kaempferol−3−o−glucoside 7 geranyl lignin−3−o−rutoside 8 Geranium rutoside 9 geranium malonyl glucoside 10 delphinidin arabinose 11 cyanidin Sophorae 12 paeoniflorin glucose 13 chlorogenic acid 14 isorhamnetin−}
^{−7−o−rutoside 15 quercetin−3−odiglucoside 16 isorhamnetin−3−o−glucoside 17 malvacin glucoside 18 geranium glucoside 19 trefoil glycoside 20 quercetin−3−o−mulberry glycoside 21 delphinidin sophoricose 22 delphinidin hexyl glucoside 23 cyanidin arabinoside)}

According to the result analysis in Table 9., 23 pigment components were detected from 12 species and hybrids of Lonicera L., including 11 flavonoid pigments and 12 anthocyanin pigments. Vinblastin, quercetin-3-o-rutoside, quercetin-3-o-glucoside, kaempferol-3-o-glucoside and paeoniflorin-glucoside are the common components in most Lonicera L. Paeoniflorin glucoside accounts for the largest proportion of anthocyanins in Lonicera L., which is the main component of fruit pigments in Lonicera L..

3.4.2. Cluster analysis of fruit pigment components of 12 species and hybrids of Lonicera L.

The 23 fruit pigments components of 12 species and hybrids of Lonicera L. were analyzed by principal component analysis (Table 10.).

Table 10

Seven principal components and its contribution rates
Principal components	Y1	Y2	Y3	Y4	Y5	Y6	Y7
Characteristic value	6.356	4.585	2.52	1.961	1.471	1.232	1.121
Contribution rate (%)	30.265	21.831	11.999	9.338	7.006	5.865	5.338
Cumulative contribution rate (%) (%) (%) (%)	30.265	52.096	64.095	73.433	80.439	86.304	91.642

Table 10. shows that the contribution rate of principal components reach 91.64%, indicating that the information contained in the principal components can fully reflect most of the information of the fruit components of Lonicera L. which can be used as the inclusion items in cluster analysis.

Cluster analysis was conducted on the fruit pigment components of 12 species and hybrids of Lonicera L. and taxonomic tree diagram was drawn (Fig. 4.).

Clustering results show that at the genetic distance of 10.0, the 12 species are divided into three groups: the first group consists of Lonicera edulisTurcz., Lonicera korolkow‘Zabelii’.×Lonicera tatarica Linn.No.8, Lonicera ruprechtiana Regel., Lonicera korolkow‘Zabelii’.×Lonicera tatarica Linn.No.2, Lonicera korolkowi ‘Zabelii’., Lonicera korolkow‘Zabelii’.×Lonicera maackii (Rupr.) Maxim., Lonicera korolkow‘Zabelii’.×Lonicera tatarica Linn.No.4, Lonicera maackii (Rupr.) Maxim., Lonicera tatarica Linn. and Lonicera ferdinandiFranch.. The second group only consists of Lonicera chrysanthaTurcz. ex Ledeb..The third group only consists of Lonicera korolkow‘Zabelii’.×Lonicera tatarica Linn.No.1..

The first group contains vinblastin, quercetin-3-o-rutoside and kaempferol-3-o-glucoside. The second group contains only four anthocyanins. In the third group, there are unique trilobatin, quercetin-3-o-sambuoside, delphinidin Sophora sugar, delphinidin hexoyl glucoside and cyanidin arabinoside in the Lonicera korolkow‘Zabelii’.×Lonicera tatarica Linn.No.1, so it was grouped into one group separately.

According to the clustering results of floral pigment components: in the first group, Lonicera korolkowi ‘Zabelii’. and Lonicera tatarica Linn. are the parents of Lonicera korolkow‘Zabelii’.×Lonicera tatarica Linn.No.2, Lonicera korolkow‘Zabelii’.×Lonicera tatarica Linn.No.4 and Lonicera korolkow‘Zabelii’.×Lonicera tatarica Linn.No.8.; Lonicera korolkowi ‘Zabelii’. and Lonicera maackii (Rupr.) Maxim. are the parents of Lonicera korolkow‘Zabelii’.×Lonicera maackii (Rupr.) Maxim. Both parents and hybrids are clustered in one group. And Lonicera korolkowi ‘Zabelii’., Lonicera ruprechtiana Regel., Lonicera tataricaLinn. and Lonicera maackii (Rupr.) Maxim. are clustered in one group, which is consistent with the morphological taxonomy.

The taxonomic relationship of pollen morphology characteristics of 13 species and hybrids of Lonicera L.

Jamzad Z and Hasani nejad M proved that surface ornamentation is an important basis for classification in this genus by studying the morphological characteristics of 29 kinds of pollen of two subgenera of Scutellaria and identifing the types of pollen surface ornamentation(Jamzad and Hasani-Nejad 2014). LiHuaicheng et al. divided 17 species of Desmodium Desv. into 4 groups according to the cluster analysis on the pollen(Huaicheng Li et al. 2018). The research results on the palynological characters of Desmodium have certain guiding significance for the classification of its subordinates.^[18]. In this paper typical characteristic indexes of 13 species and hybrids of Lonicera L. are selected for cluster analysis.

The results that both parents and hybrids, Lonicera korolkowi ‘Zabelii’., Lonicera ruprechtiana Regel., Lonicera tatarica Linn. and Lonicera chrysanthaTurcz. ex Ledeb as well as Lonicera ferdinandiFranch. and Lonicera edulisTurcz. are clustered in one group are consistent with morphological taxonomy, indicating that the results of pollen morphological classification are highly consistent with the morphological classification of Lonicera L. and providing a palynological basis for the classification of Lonicera L.

The taxonomic relationship of Chromosome karyotype characteristics of 13 species and hybrids of Lonicera L.

Chuanmei Xu et al. clarified the genetic relationship of 10 species of Carya cathayensis according to their karyotypes(Chuanmei Xu 2017). In this paper,8 typical characters of chromosome morphology of 13 species and hybrids Lonicera L. were selected for cluster analysis.

The results that both parents and hybrids, Lonicera maackii (Rupr.) Maxim. and Lonicera tatarinowiiMaxim. as well as Lonicera tatarica Linn. and Lonicera ruprechtiana Regel. belonged to the category of hollow branchlets are clustered in one group are consistent with morphological taxonomy, showing that the chromosome morphological classification is partly consistent with the morphological classification of Lonicera L., which indicates that the results of this study are scientific.

The taxonomic relationship of floral pigment components of 13 species and hybrids of Lonicera L.

Masaji et al. divided the genetic relationships of 9 species of Curcuma into two categories according to 11 kinds of anthocyanins detected from them(Koshioka et al. 2015). In this paper, 13 species and hybrids Lonicera L. are divided into three groups according to 26 anthocyanins.

The results that both parents and hybrids as well as Lonicera korolkowi ‘ZabTelii’., Lonicera ruprechtiana Regel. and Lonicera tatarica Linn. are clustered in one group are partly consistent with the morphological taxonomy of Lonicera L., indicating that the floral pigment components has a certain scientific value for the classification of Lonicera L..

The taxonomic relationship of fruit pigment components of 12 species and hybrids of Lonicera L.

Wang Y et al. divided 44 firework trees into three types according to the anthocyanins which identified from the plants(Wang et al. 2018).In this paper, 21 anthocyanins identified from 12 species of Lonicera L. were clustered,12 species and hybrids of Lonicera L. are divided into three groups. Both parents and hybrids are clustered in one group. And Lonicera korolkowi ‘Zabelii’., Lonicera ruprechtiana Regel., Lonicera tataricaLinn. and Lonicera maackii (Rupr.) Maxim. are clustered in one group, which is consistent with the morphological taxonomy.

In morphological taxonomy, the berries of Lonicera edulisTurcz. are long ellipse in shape and deep blue with a coat of powder in color. The berries of the other 11 species (varieties) Lonicera L. are spherical in shape and red in color. Although there are great differences in morphology, Lonicera edulisTurcz. is divided into corresponding groups during clustering in this paper. Ying Zhu et al. believed that Lonicera edulisTurcz. belongs to the blue-fruited subgroup of Lonicera L., which is far away from other Lonicera L.. This view explains one phenomenon that Lonicera edulisTurcz. belongs to Lonicera L., but far away from the other Lonicera L. in the clustering results(Ying Zhu et al. 2009).

In this paper, 13 species and hybrids of Lonicera L. were clustered according to pollen morphology characteristics, chromosome karyotype characteristics, floral pigment and fruit pigment components. It is found that the cluster results that Lonicera korolkowi ‘Zabelii’., Lonicera ruprechtiana Regel., Lonicera tatarica Linn. and Lonicera chrysanthaTurcz. ex Ledeb. as well as Lonicera ferdinandiFranch. and Lonicera edulisTurcz. are clustered in one group are most consistent with morphological taxonomy. Five hybrids such as Lonicera korolkow‘Zabelii’.×Lonicera maackii (Rupr.) Maxim. and Lonicera korolkow‘Zabelii’.×Lonicera tatarica Linn.No.8 all can basically be clustered with their parents. Among the four clusters, Lonicera ruprechtiana Regel and Lonicera tatarica Linn. are always clustered together, which is consistent with morphological taxonomy, indicating that pollen morphology characteristics, chromosome karyotype characteristics, floral pigments and fruit pigments components have certain scientific significance for the study of systematic classification and genetic relationship of Lonicera L. However, it is found that the classification results of Lonicera L. at these four levels are not completely consistent, there being some differences, indicating that the evolution of Lonicera L. at the levels of secondary metabolites, palynology and cell biology is not very consistent.

Acknowledgements

The authors are very grateful to College of horticulture and gardening of Northeast Agricultural University for funded this study.

Author Contributions

Da LongLi: Determination of flowers and fruits pigment components

Xiaoou Zhai: Observation and measurement of pollen morphological

Li Fang: Sampling and photographing of chromosome karyotype

Liwei Sun and Xing Ji: Data curation

Sijia Zhang: Image processing

Yujiao Zhang: Cluster analysis

Liu Huimin*: Writing - original draft

Funding

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The datasets generated during and/or analyzed during the current study are

available in supplementary table fle.

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Yes, we agreed and give consent for participation.

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Authors give consent for publication of this manuscript.

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The authors declare no conficts of interest that are relevant to the content of this article.

Author details

¹Northeast Agricultural University, Harbin, China. ² Heilongjiang Forest Botanical Garden, Harbin, China. ³ Qiqihar University, Qiqihar, China. ⁴Heilongjiang Academy of Forestry, Harbin, China. ⁵ Shandong Shouguang testing group co ltd, Shouguang, China. 6Agricultural comprehensive administrative law enforcement brigade of Melis Daur Nationality District, Qiqihar, China

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PlateI.jpeg
Pollen morphology and exine ornamentation characteristics of 9 species of Lonicera L.（A: Lonicera tatarica Linn. B: Lonicera tatarinowii Maxim. C:Lonicera korolkowi ‘Zabelii’.×Lonicera maackii (Rupr.) Maxim.; D: Lonicera tatarinowii Maxim.; E: Lonicera korolkowi ‘Zabelii’.×Lonicera maackii (Rupr.) Maxim.; F: Lonicera korolkowi ‘Zabelii’.×Lonicera tatarica Linn.No.4;G: Lonicera korolkowi ‘Zabelii’.×Lonicera tatarica Linn.No.4;H: Lonicera korolkowi ‘Zabelii’.×Lonicera tatarica Linn.No.8; I: Lonicera korolkowi ‘Zabelii’.×Lonicera tatarica Linn.No.2）
PlateII.jpeg
Pollen group view under SEM of three species of Lonicera L.（A: Lonicera maackii (Rupr.) Maxim.; B: Lonicera ferdinandi Franch.; C: Lonicera korolkowi ‘Zabelii’.）
PlateIII.jpeg
Chromosome characteristics of three species of 3species Lonicera L.(A: Lonicera korolkowi ‘Zabelii’.; B: Lonicera tatarinowiiMaxim.; C: Lonicera korolkowi ‘Zabelii’.)

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A taxonomic study on pollen morphology characteristics, chromosome karyotype characteristics, floral and fruit pigment compositions of 13 species and hybrids of Lonicera L.

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Abstract

Figures

1. Introduction

2. Materials And Methods

2.1. Experimental materials

2.2. Experimental method

3. Results

3.1. Taxonomic relationship of 13 species and hybrids of Lonicera L. based on pollen characteristics.

3.1.1. Pollen morphological characteristics and value assignments of 13 species and hybrids of Lonicera L.

3.1.2. Cluster analysis of pollen morphological characteristics of 13 species and hybrids of Lonicera L.

3.2. Taxonomic relationship of 13 species and hybrids of Lonicera L. based on chromosome karyotype characteristics.

3.2.1. Chromosome karyotype characteristics and value assignments of 13 species and hybrids of Lonicera L.

3.2.2. Cluster analysis of chromosome karyotype characteristics of 13 species and hybrids of Lonicera L.

3.3. Taxonomic relationship of 13 species and hybrids of Lonicera L. based on floral pigment compositions.

3.3.1. Floral pigment compositions analysis and value assignments of 13 species and hybrids of Lonicera L.

3.3.2. Cluster analysis of floral pigment components of 13 species and hybrids of Lonicera L.

3.4. Taxonomic relationship of 12 species and hybrids of Lonicera L. based on fruit pigment components.

3.4.1. Fruit pigment components analysis and value assignments of 12 species and hybrids of Lonicera L.

3.4.2. Cluster analysis of fruit pigment components of 12 species and hybrids of Lonicera L.

4. Discussion And Conclusion

Declarations

References

Supplementary Files

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