NL2032854B1 - Artificial crossbreeding method for tea plants - Google Patents

Abstract

Disclosed is an artificial crossbreeding method for tea plants and belongs to the field of 5 crossbreeding for plants. According to an objective of breeding with high quality, strong fragrance, high amino acids, high yield and high resistance, resources are collected and matched for combination. In order to solve the problem that backbone parents are highly concentrated, in the present invention, 1735 domestic and foreign resources had been consecutively collected and preserved from 1976 to 2019 and had been widely combined, and 86 resources were matched 10 and combined. Through researches on trait characteristics, fruiting rate, pollen viability and times and durations of full-bloom stages of hybrid parents, genetic relationship between the hybrid parents and the like, 12 varieties suitable for serving as parents are proposed, wherein 9 varieties are more suitable for serving as female parents; and 3 varieties are more suitable for serving as male parents. In the aspect of an artificial crossbreeding method, in a pollination method, a 15 combination of single-crossing, pollination with mixed pollens from multiple male parents and free pollination is employed; a method ofa desiccant plus a low temperature to preserve and transport pollens is employed; methods of pollination immediately after emasculation of flowers in a bud white part appearing stage and the like are proposed; and a comprehensive evaluation method for hybrid tea seedlings in F1 generation is proposed. Through directional selection, 10 new hybrid 20 varieties are selected and registered, 18 new varieties enter expanding propagation to be registered, and 77 new lines, 302 fine new hybrid varieties in F1 generation and 955 fine individual plants are identified.

Description

ARTIFICIAL CROSSBREEDING METHOD FOR TEA PLANTS

Technical Field

The present invention relates to the field of crossbreeding of plants, in particular to an artificial hybridization technology and a breeding method for tea plants.

Background

China is one of countries where tea plants are planted first in the world and is also the country that has the largest tea garden area in the world and is largest in the yield of teas, accounting for 62.5% of the tea garden area in the world and 45% of the yield.

China is also the most important tea export trading country in the world, ranking second and third in the world; and good tea export trade and domestic sale promote stable development of the tea industry. “Farmers focus on varieties”. A tea plant variety is the most basic and most important production resources of the teas; a fine tea plant variety plays a very significant role in improving the yield and the quality of the teas, increasing labor productivity and improving the stress resistance and is the basis of achieving sustainable development of the tea industry.

There are many ways of germplasm innovation and variety selective breeding for modern crops, such as variety introduction, selective breeding, mutation breeding, crossbreeding, ploidy breeding and biotechnology breeding, wherein artificial crossbreeding is the most economical and most effective means for germplasm innovation and variety selective breeding of the tea plants and is also an important basis of researching trait heredity laws of the tea plants.

Strengthening innovation of an artificial crossbreeding technology of the tea plants and solving the technical bottleneck problems of high concentration in backbone parents, long breeding cycle and the like of traditional artificial crossbreeding are the key of rapidly obtaining a new ground-breaking germplasm. 1

Summary 1 A tea plant breeding objective of the present invention takes a traditional high yield as a core, and newer (more) objectives for high quality, strong fragrance, high amino acids, high yield and high resistance are proposed. 2 In order to solve the problem of high concentration in backbone parents of artificial crossbreeding, in the present invention, 1735 resources including 3 species of Camellia tachangensis, Camellia taliensis and tea of the Genus Camellia Sect. Camellia in the family Camellia and 2 varieties of Baimao Tea and Assam Tea from home and abroad had been consecutively collected and preserved from 1976 to 2019, which provides a rich germplasm gene bank for artificial crossbreeding of tea plants. 3 Screening of hybrid parents comprises following several researches: 3.1 prominent one or more of traits such as quality, yield and resistance; 3.2 fruiting rate; 3.3 strength of pollen viability; 3.4 times and durations of full-bloom stages; 3.5 genetic relationship between main resources. 4 Artificial hybrid combination and matching 4.1 Combination principle: artificial hybrid combination and matching require to make combination according to the breeding objective and the principles of prominent features and complementary traits of male and female parents. 4.2 Important parents: corresponding 12 varieties suitable for serving as female parents and male parents are proposed, wherein 9 varieties are more suitable for serving as female parents; and 3 varieties are more suitable for serving as male parents (Table 1).

Table 1 Highly Recommended Parents

Female Jinxuan, Tieguanyin, Fuding White Tea, Yabukita, C.sinensis cv.Bixiangzao, parent Wuniuzao, C.sinensis cv.Baihaozao, Anhui No. 1, and Zhuyeqi i” Yunkang No. 10, Baojing Huangjin tea No. 1, and Yingde Black Tea No. 9 2

4.3 Combination method: there are two kinds: combination breeding and transgression breeding. In this research, the two are combined together, so that fine new clonal varieties can be selected, and double hybrid clonal sexual varieties can further be selected and bred.

Table 2: Artificial Hybrid Varieties of Tea Plant Selected and Bred in Hunan Province.

Complete unit: Tea Research Institute of Hunan Province.

Variety name / Selection Female

Number {selection code Male parent Year mode parent number)

C.sinensis == Artificial Fuding White 1 cv.Bixiangzao (79-30- Yunnan Large-leaf Tea 1993 ) hybridization Tea / Artificial Fuding White 2 Mingfeng (79-30-2) Yunnan Large-leaf Tea 1994 hybridization Tea

C.sinensis cv.Fuhao Artificial Fuding White / 3 Large-leaf Changqing 1996 (79-38-9) hybridization Tea

Yunnan Big

Hunan Black Tea No. Artificial 4 Co Leaf Species Fuding White Tea 1998 1 (79-6-7) hybridization

Tea

C.sinensis

Hunan Black Tea No. Artificial 3 Co 5 — cv. Fujian- Preferred hybridization 2003 2 (79-10-04) hybridization oo shuixian 6 Yusun (81-8-65) Mixed pollens of excellent 2009 varieties of Fuding White

Artificial

Yabukita Tea, Zhuyeqi, Xiangbolv No. hybridization 7 Yulv (81-8-30) 2, Longjing No. 43 and the 2010 like

Mixed pollens of 4 varieties

Xiaoxiang No. 1 (80- Artificial / (strains) of Sichuan Gulin 8 — Xiangbolv oo / 2011 53-34) hybridization niupicha, Gaoyaqi, Fuyun

No. 7 and Yunda 72-04 / Yunnan Big

Xiangchayan No. 2 Artificial / / / 9 — Leaf Species Fuding White Tea 2019 (79-86-16) hybridization

Tea 3

10 Xiangchayan No. 6 Artificial Zhuyeqi Vietnam Big Leaf Species oet (79-28-86) hybridization Tea cation 4.4 Directional selection and breeding: from performance of F: generation, 2 double hybrid clonal combinations with strong hybrid superiority, 955 dominant individual plants, 302 new strains and 77 new lines (wherein 18 new lines may apply for registration} are screened, and 10 artificial hybrid varieties have been approved (registered) (Table 2).

Detailed Description

Specific implementations of the present invention will be introduced below. The present invention can be implemented in many other ways different from those described here, and those skilled in the art can make similar improvements without departing from the essence of the present invention. Therefore, the present invention is not limited by the specific implementations disclosed below.

Unless defined otherwise, all technical and scientific terms used herein have the same meanings as commonly understood by those skilled in the art to which the present invention belongs. The terms used in the specification of the present invention herein are for the purpose of describing particular embodiments only and are not intended to be limit the present invention. 1 Unless specified explicitly, the technical means in the embodiments are means and methods explored by those skilled in this research subject. 2 Research on genetic relationship. The present invention uses 33 pairs of polymorphic SSR primers to analyse the genetic similarity and genetic distances among collected domestic and foreign resources and selects and combines 56 hybrid combinations according to above results, variety features and the combination principle (Table 3). 4

Table 3 Hybrid Combination and Matching ~~ Femaleparet ~~ Maleparent(?) Similarity Breeding

Number EE (£) Variety name Trait (advantage) coefficient objective

Strong

Earliness in germination fragrance, stage, much more hairs, many hairs, 1 Fuding White Tea” high yield, strong 0.58 cold resistance, wide resistance, adaptability earliness in germination

Extremely early budding, Early large bud density, suitability budding, 2 Wuniuzao for processing Green Tea, 0.47 specificity particularly flat-shaped for Green famous Green Tea Tea

The processed Green Tea is full of white hairs, high in High 3 C.sinensis cvBaihaozao’ quality and strong in 0.56 quality, high resistance, particularly resistance drought resistance

Strong

Anhui No. 1° The yield is high, fragrance, 4 (high polyphenol, Vingde Black Tea No. 9 particularly, the processed 0.53 high quality strong fragrance) Black Tea is excellent in specificity quality and full of gold hairs for Black

Tea

Strong

High yield, many hairs, wide fragrance, suitability for preparation, high quality, particularly the processed suitability

Baimao Tea No. 2° 0.50

Green Tea and Oolong Tea for having strong and long floral processing fragrances many tea types

Good

The processed Black Tea tenderness- and Green Tea are high in keeping 6 Yunkang No. 10° quality, strong and long in 0.58 ability, floral fragrance and strong strong in resistance and cutting resistance, reproductive capacity reproductiv e capacity 5

Strong cold resistance, processing / suitability for processing both of

Gaoyadi 7 / Green Tea and Black Tea, 0.55 Black Tea (Zhuyegi No. 9) and strong cutting and Green reproductive capacity Tea

Much more hairs, suitability for processing Green Tea, Strong 8 Yulv' finished tea having the 0.60 fragrance, characteristic of ‘three Green Tea green” . i Green Tea,

High yield and strong top / 9 Yusun 0.61 mechanical growth vigour / plucking

Suitability for processing

Strong buds, the processed single bud

Black Tea and Green Tea / Co . / teas (such

Taoyuan Bigleaf Tea are high in quality, strong in 0.58 / as Silver drought resistance and cold / Needle Tea) resistance and Dark

Tea, strong resistance

Strong fragrance,

Long internode, elegant / / mechanical i fragrance with fragrance of /

Jinxuan / plucking, 11 . magnolia flowers, strong 0.59 (Tai Tea No. 12) A / double flowering and fruiting ability, . . hybrid strong resistance clonal variety

High amino , / Japan leading green tea / 12 Yabukita variety / 0.62 acids, large variety, strong fragrance bud density

Extremely early budding, / / large bud density, suitability Large bud

Fuding White . . . 13 Tea Wuniuzao for processing Green Tea, 0.54 density, ea / particularly flat-shaped Green Tea (Early budding, famous Green Tea much more / / processed Green Tea has Strong hairs, wide : fragrance of orchids and is fragrance, 14 adaptability) Shuchazao / 0.58 strong in drought (cold) cold resistance, particularly resistance 6 strong in ate frost resisting ability in early spring

The processed Black Tea and Green Tea are high in / / Strong . quality, strong and long in

Yunkang No. 10 0.59 fragrance, floral fragrance and strong / Black Tea in resistance and cutting reproductive capacity

Extremely early germination . . Strong stage, high amino acid fragrance, 16 Baojing Huangjin tea No. 1 content, wide suitability for 0.63 / / / multiple tea preparation, good quality of types processed teas buds and leaves are strong in fertility and tendemess- Strong keeping ability, wide in adaptability, 17 Zhuyegi’ ep 9 y 0.59 P Y suitability for preparation multiple tea and strong in resistance and types adaptability / LE Early

Much more hairs, suitability / 18 Long-leave Baihao 0.52 budding, for processing Green Tea

Green Tea

Strong — High tea polyphenol resistance 19 Xiaoxianghong 21-3 0.54 content, strong resistance and Black

Tea

Long intemode, strong stress / . fruiting ability, high survival resistance,

Hubei Tea No. 1 J 0.54 / rate of transplanting, strong mechanical stress resistance plucking

Strong

High olyphenol, stron fragrance, 21 Anhui No. 1” ah pobp ¢ 0.58 a0 . fragrance high quality,

Green Tea

Strong

Strong fragrance, high fragrance, 22 Huangjin tea No. 2 / / 0.62 / / amino acids high quality,

Green Tea / processing

Strong cold resistance, both of . suitability for processing

Gaoyaqi Black Tea 23 / Green Tea and Black Tea, 0.58 (Zhuyegi No, 9) / A and Green strong cutting reproductive

Tea, cold capacity / resistance 7 large bud

Japan leading green tea density, 24 Yabukita variety . 0.71 variety, strong fragrance cold resistance

The processed Black Tea and Green Tea are high in ] / / High . quality, strong and long in

Yunkang No. 10 0.51 polyphenol, floral fragrance and strong / i Black Tea in resistance and cutting

Zhuyeqi’ reproductive capacity (Strong Stress tenderness- The processed Green Tea is resistance, keeping ability full of white hairs, high in processing 26 and adaptability, C.sinensis cv. Baihaozao’ quality and strong in 0.62 both of wide suitability resistance, particularly Black Tea for preparation) drought resistance and Green

Tea

Long internode, strong Stress / . fruiting ability, high survival resistance, 27 Hubei Tea No. 1 i 0.56 rate of transplanting, strong mechanical stress resistance plucking

Stress resistance,

Flavor of big leaf species, processing 28 Assam good quality of processed 0.49 both of

Black Tea Black Tea and Green

Tea

High yield, many hairs, wide suitability for preparation,

Strong

Yabukita variety . . particularly the processed 29 Baimao Tea No. 2 0.56 fragrance, (Japan leading Green Tea and Oolong Tea / Green Tea green tea variety, having strong and long floral strong fragrance, fragrances cold resistance) Extremely early budding, large bud density, suitability Early

Wuniuzao for processing Green Tea, 0.57 budding, particularly flat-shaped Green Tea famous Green Tea

The processed Green Tea is full of white hairs, high in More hairs, 31 C.sinensis cv.Baihaozao' quality and strong in 0.56 early resistance, particularly budding drought resistance 8

. processing

Strong cold resistance, both of

Rl suitability for processing

Gaoyadi Black Tea 32 Green Tea and Black Tea, 0.59 (Zhuyegi No. 9) / and Green and strong cutting / Tea, cold reproductive capacity / resistance

Strong

Flavor of big leaf species, fragrance, 33 Assam good quality of processed 0.45 Black Tea,

Black Tea stress resistance

The yield is high, Strong / particularly, the processed fragrance, 34 Yingde Black Tea No. 9 0.58

Black Tea is excellent in more hairs, quality and full of gold hairs Black Tea i Strong / / Japan leading green tea

Yabukita variety . 0.65 fragrance, variety, strong fragrance

Green Tea

Extremely early germination . . Stress stage, high amino acid / / resistance, 36 Baojing Huangjin tea No. 1 content, wide suitability for 0,51 A / : multiple tea

Jinxuan preparation, good quality of t ypes (Long internode, processed teas elegant / Strong i . Strong fragrance, high 37 fragrance with Huangjin tea No. 2 / . 0.58 fragrance, amino acids fragrance of Green Tea magnolia | More hairs,

Much more hairs, suitability flowers, strong processing . for processing Green Tea, resistance) both of 38 Yulv” finished tea having the 0.51

Co Black Tea characteristic of ‘three and Green green”

Tea

High yield, good quality of High yield, 39 Mingfeng ah y 3 a y 0.45 oy processed Green Tea Green Tea

Extremely early budding, Strong large bud density, suitability fragrance,

Wouniuzao for processing Green Tea, 0.47 early particularly flat-shaped budding, famous Green Tea Green Tea

High yield,

Wide suitability for strong 41 C.sinensis cv.Bixiangzao preparation, strong 0.54 fragrance, adaptability, high yield multiple tea types 9 processed Green Tea has fragrance of orchids and is Specific strong in drought (cold fragrance 42 Shuchazao’ 9 ght (cold) 0.58 9 resistance, particularly type, cold strong in ate frost resisting resistance ability in early spring

Mechanical / plucking,

Long internode, strong . A A double ] . fruiting ability, high survival / 43 Hubei Tea No. 1 / 0.58 hybrid rate of transplanting, strong clone, stress resistance stress resistance

The processed Black Tea and Green Tea are high in Strong uality, strong and long in fragrance, 44 Yunkang No. 10° Way 9 9 0.57 g floral fragrance and strong stress in resistance and cutting resistance reproductive capacity

Strong

High olyphenol, stron fragrance, 45 Anhui No. 1* gh pop 9 0.59 9 fragrance stress resistance

Extremely early budding, large bud density, suitability Early 46 Wuniuzao for processing Green Tea, 0.38 budding, particularly flat-shaped Green Tea famous Green Tea

C.sinensis High yield, cv.Baihaozao’ Wide suitability for strong 47 (The processed C.sinensis cv.Bixiangzao preparation, strong 0.47 fragrance,

Green Tea is full adaptability, high yield multiple tea of white hairs, types high in quality A budding density is large, . Strong and strong in processed Oolong Tea also . . we fragrance, 48 resistance, Tieguanyin has a name of “fragrance 0.54 high vield i ield, particularly soaring”, and the yield is MY

Green Tea drought high resistance) Early

Long internode, strong budding, / . fruiting ability, high survival stress 49 Hubei Tea No. 1 / 0.47 / rate of transplanting, strong resistance, stress resistance mechanical plucking

Suitability for processing

Cold . Green Tea, strong cold 50 Xinyang No. 10 / 0.54 resistance, resistance and drought

Green Tea resistance i Strong

High polyphenol, strong 51 Anhui No. 1* 0.56 fragrance, fragrance

Green Tea

High yield, good quality of High yield, 52 Mingfeng any 3 qua ty 0.44 oy processed Green Tea Green Tea / Early

Extremely early germination / budding, stage, high amino acid stress 53 Baojing Huangjin tea No. 1 content, wide suitability for 0.52 / resistance, preparation, good quality of multiple tea processed teas types

A Strong ‚ Strong fragrance, high 54 Huangjin tea No. 2 . / 0.57 fragrance, amino acids

Green Tea

Strong cold resistance, . Stress . suitability for processing .

Gaoyaqi resistance, 55 / Green Tea and Black Tea, 0.51 / {Zhuyegi No. 9) / multiple tea and strong cutting . . types reproductive capacity

Strong fragrance, . } Japan leading green tea 56 Yabukita variety 0.56 stress variety, strong fragrance . resistance,

Green Tea

Note: “*” represents national fine variety 3 Research on fertility of tea plants. Strength and existence of the fertility of tea plant varieties are primary conditions of selecting parents. The fruiting rate (fertility) of a female parent is researched through wide combination, a pollen germination rate is determined, and the strength of pollen viability is researched. As a result, the fertility of the tea plants is divided into three types: the first type is that the parent is normal in gender function and thus can serve as both the male parent and the female parent and can be more suitable for serving as the female parent; the second type is that the parent is low in fruiting rate and strong in pollen viability and is more suitable for serving as the male parent; and the third type is that the parent is in infertility. 11

4 Pollen storage method.

The pollen viability of different varieties under different storage conditions is determined by using an in vitro pollen germination method.

Results show that under the storage condition of low temperature plus a desiccant, preservation of the pollen viability is facilitated.

Thus, the problem that the flowering stages of the male parent and the female parent do not meet each other is solved.

5 Hybridization pollination technology.

Hybridization pollination technologies of artificially emasculated pollination and bagging, pollination and bagging without emasculation, artificial pollination without emasculation and bagging, different flowering ages and the like are researched.

Results indicate that crossbreeding with a trait heredity research as a major purpose should employ emasculated bagging; hybridization pollination requires to be conducted in a bud white part exposure stage; and tea flowers require to be pollinated immediately after being emasculated, and then a high fruiting rate can be provided.

6 Crossing mode.

The present invention employs two methods of single crossing and mixed crossing (pollination with mixed pollens from multiple male parents). 7 varieties such as C.sinensis cv.Bixiangzao are selected and bred by using the single- crossing pollination mode; and 3 varieties of Xiaoxiang No. 1, Yulv and Yusun have been selected and bred by using pollination with the mixed pollens from the multiple male parents.

7 Tea seed harvesting and storage.

Artificial hybridization pollination is conducted from early October to mid-to-late November in a first year, tea seeds are harvested in about one week before and after Frost's descent in a next year, and a fruiting rate is calculated.

The harvested tea seeds are stored in a shade after being tested; tea seeds required to be sown in spring in the next year in Jiangbei Distinct may be moisturized and stored with fine sands; and tea production regions in Jiangnan region, South China and Southeast China best employs winter sowing in a current year.

8 Sowing and seedling raising.

Before sowing and seedling raising of the tea seeds, the tea seeds require to be soaked into 3% carbendazim (with a content of 50%) solution for 3 days with replacing water once a day and are washed with clean water and dried

12 in the air for sowing finally; Seedling raising employs single-seed drilling with a row space of 10 cm, a seed space of 3 cm and shallow seeding (3-5 cm). After sowing, 2-3 cm straws are laid, water should be so poured and sprayed that the part at about 5 cm depth below the earth surface is wet, and then a mulching film covers the straws (which may improve ground temperature and preserve moisture). When the temperature reaches about 20°C in March-April, the film is uncovered. 9 Determination of emergence rate and seedling rate. Emergence rates are surveyed in late April and early May once every 15 d; and if results of consecutive 2 surveys are the same, a seedling rate may be calculated. When tea seedlings stop growing in late autumn and early winter, the seedling rate is surveyed, and surveys are generally conducted in November in the tea regions in the Jiangnan region. Standards of clonal tea seedlings refer to Seedlings For Tea Plants (GB 11767—2003) and divided into grade | and grade II (Table 4); and the clonal tea seedlings not reaching the standards are not counted in the seedling rate.

Table 4 Quality Standard For Seedlings

Grade Seedling age Seedling height Stem diameter (mm) (cm)

Grade | —Full age 230 23.0

Grade II —Full age 220 22.0 10 Identification on fine plants in F4 generation.

From 2009 to 2017, 14525 plants in artificial F4 hybrids reaching the seedling standard were planted according to 140 cmx33 cm single-row single plant; 9 selective nurseries were built year by year; and fine plants were selected and identified for consecutive years. 10.1 Identification on growth vigour of tea seedlings in Fy generation. The growth vigour of one-year-old seedlings of the tea plants in F+ generation is surveyed; a plant height, a stem diameter, a number of leaves and a branching number of each individual plant are determined; original data is made nondimensional; a trait weight scheme is 13 designed as follows: plant height: stem diameter: branching number (1:1:1); scores are counted plant by plant; and the growth vigour of the tea seedlings is measured according to a weighted composite score value. 10.2 Selection and identification on fine plants in F1 generation. Under the same environment conditions and cultivation conditions, artificial selection is conducted according to a breeding objective.

The present invention employs a method combining independent elimination selection with trait weighting selection. 10.2.1 Selection of plant type and growth vigour. Colors of buds and leaves, sizes of leaves, leaf shapes, quality of the leaves, glossiness, lengths of internodes and the growth vigour are visually measured. 10.2.2 Identification on phenological stage of vegetative buds. With Fuding White

Tea (of early variety) as a control, the vegetative buds are divided into extremely early budding, early budding, medium budding and late budding. 10.2.3 Identification on suitability for processing tea types. A trichloromethane fermentation test is conducted, and the fermentability is judged into three degrades, based on which, the teas are approximately divided into three types of being suitable for processing Green Tea, being suitable for processing Black Tea and being suitable for processing both Green Tea and Black Tea. 10.2.4 Identification on resistance. Field natural identification is conducted on high temperature resistance, drought resistance, cold resistance, disease resistance (mainly anthracnose resistance) and the like of tested individual plants. The drought resistance/cold resistance is divided into strong, relatively strong, medium and weak; and the weak tested individual plants are not included. The tea anthracnose resistance is divided into resistance, medium resistance, sensitivity and high sensitivity; and the sensitive or high sensitive individual plants are not included. The individual plants with weak resistance are all independently eliminated. 14

10.2.5 Yield identification. Shoot and leaf weights from first to third shaping and pruning of primarily selected individual plants are determined to serve as early yields of fine individual plants.

A decision is made according to the above 5 trait weights. For consecutive 9 years, 955 fine individual plants are selected from 14525 individual plants in artificial F1 hybrids, and an average optimum selection rate is 6.57% (Table 5).

Table 5 Hybrid F1 Fine Plant Selection Rate from 2009 to 2018

Sowi ber and Number of fi

Wing NUMBET and emergence Number of fine individual plants um er of ine number stains

Number rrr, rere || eee

Cross . . First ‚ Selective of Sowing Emergence Emergence . Second

Ing ‚ optimum ‚ . . . nursery combinat number number rate . optimum Fine plant Inclusion Inclusio year ‚ selection . . ions selection selection number n rate (rain) plant 1 number NET rate (%) (plant) {%) g p 6 (plant) (plant) 2009 DE ction 19/38 613 165 26.92 18 18 10.91 7 4.24 1 2010 een 68/68 4849 3424 70.61 1695 344 10.05 159 4.64 1 2011 DE 41/41 4607 2843 61.71 1211 138 4.85 74 2.6 1 2012 VIET 45/45 2944 1685 57.24 1267 114 677 62 3.68 1 lecti T 2013 22€CHON 7/27 1569 1088 69.34 567 61 5.61 obe , (VY Fs selected 2014 Selection 43/43 3730 2264 60.7 1509 160 7.07 Tobe / (VI) F1 selected

Selection To be 2015 33/36 4544 2058 45.29 652 92 4.47 (VI F4 3/3 05 selected /

Selection To be 201 14/2 1 2 40.27 9 2 .89 6 (VRB Fy 125 788 720 390 8 3 selected /

Selection 2017 14/25 732 278 37.98 / / / / / (IX) F1 2018 -- -/26 / / / / / / / /

Average 3.79

Total 304+/374 25376 14525 7309 955 6.57 302

10.3 Comparison on cutting reproductive capacity of fine individual plants in F4 generation. Totally 436 4-5-year-old fine individual plants are screened according to the above method for cutting in autumn (mid-to-late September); in a next year, a survival rate is surveyed in late April, the growth vigour is surveyed in November, and plant heights, stem diameters, branching numbers and numbers of leaves are determined; and referring to GB 11767-2003 standards, a qualified seedling percentage is calculated with numbers of seedlings reaching the grade | and the grade Il. 10.4 Comprehensive identification on fine strains. Cutting seedlings of 302 individual plants are selected from the above 436 cutting fine individual plants for field planting; and the biological characteristics are observed, and the resistance, the quality and the yield are primarily identified. 10.5 Variety comparative test. 85 plants are selected from the above 302 fine individual plants to assign a variety comparative test. The variety comparative test employs random arrangement with three replications, wherein a plot area is 0.02 mu (9 m x 1.5 m), and a planting distance is 1.5 m x 0.33 m; and a test method refers to the national regional trial requirements, and the quality, the yield and the resistance are systematically identified. 8 high-amino-acid (>5%) lines, 13 high-quality and strong- fragrance new lines and 25 high-yield new lines are selected and bred, wherein 4 new lines are high in quality and fragrance and also high in yield; and totally 42 special new lines are selected and bred (Table 6, Table 7, Table 8, Table 9).

Table 6 High-amino-acid New Lines ee Ser Combination es yohe oo acid oo ber (%) nol (%) ~~ Yunnan black a eavexXiangfei 36.85 14.29 7.58 4.10 cui 16

2012-6- TieguanyinxBa 2 ojing Huangjin 37.42 19.95 6.41 3.89 28-7 tea No. 1 2012-6- TieguanyinxFu 3 20.3 ding White Tea 36.28 17.00 6.26 3.95

C.sinensis cv.Baihaozaox 4 2912685 Baojing 38.95 19.80 6.14 3.52

Huangjin tea

No. 1

C.sinensis 2012-6- cv.Bixiangzaox 38.64 21.67 6.11 4.13 33-3 ,

Longjing 43 6 2012-8- YingshuangxTi 2445 14.79 6.09 3.66 56-6 eguanyin 2012-6- Fuding White 7 0.4 TeaxShuchaza 35.59 16.88 6.09 4.00 0

Fuding White 8 2012-6- TeaxAnhui No. 38.30 19.87 6.05 3.87 37-6 4

Baojing 9 Huangjin 35.66 18.19 5.84 3.68 tea No. 1 17

Table 7 Examination on High-Quality and Strong-Fragrance New Lines for Green Tea

Sequenc Interior quality . Total

Serial number Lo ee Total . Combination Shape scor e number of line . rank

Tea liquor color Fragrance Taste infused leaf e

Freshn ess, heavin Yellow

Bit tight, Durable faint

Baxian TeaxHuangjin tea No. Tender green, ess, green, 1 2012-6-62-1 bright 93.0 95.0 chestnut 94.5 96.0 94.0 94.58 1 2 bright sweetn bright, neat green fragrance ess and even after taste

Bright Yellow , hai . green, green, haw Faint fragrance, Freshn

C.sinensis appearing Yellow green, bright, not 2 2012-6-12-15 93.5 94.0 fresh 95.0 ess and 96.0 92.5 9458 2 cv.BixiangzaoxTieguanyin bright sufficiently ‚ sharpness mellow

Relatively neat and tight even

Relatively . Freshn Yellow bright Tender green, Tender 3 2012-8-4 Hubei Tea No. 1 93.0 95.0 95.0 ess and 945 green, 93.0 94.15 3 green, bright chestnut . . ‚ mellow bright, relatively fragrance with relatively tight, 18 having flower neat and hairs fragrance even

Freshn

Relatively ess, Yellow bright mellow green,

Bit strong faint

C.sinensis green, Tender green, , bright, 4 2012-6-67-3 93.0 95.0 chestnut 93.0 96.0 92.5 94.05 4 cv.BaihaozaoxAnhui No, 1 relatively bright sweetn relatively fragrance tight, hair ess neat and appearing after even taste

Bit bright

Yellow green, green, green with Durable faint Freshn

TieguanyinxBaojing Huangjin Tender green, bright, 2012-6-28-5 bit 91.5 96.0 chestnut 95.0 ess and 94.5 93.0 93.88 5 tea No. 1 bright relatively brightness, fragrance mellow nest and having even hairs

Green with moist bit, relatively Strong, Yellow bright durable, fresh Freshn green, . ZhuyeqixBaojing ~~ Huangjin Tender green, 6 2012-6-52-1 green, 92.0 96.0 and mellow 95.0 essand 94.0 relatively 93.0 93.85 6 tea No. 1 bright relatively flower mellow bright, neat tight, fragrance and even having hairs 19

Green with bit moist,

Yellow relatively green,

C.sinensis bright Durable tender Freshn

Yellow green, bright, 7 2012-6-9-13 cv.Bixiangzaoxiongjing Long- green, 91.5 94.5 chestnut 95.0 essand 94.5 93.0 93.73 7 bright relatively leave relatively fragrance mellow neat and tight, even having hairs

Yellow green,

Tight, Durable tender Freshn

Tender green, bright, 8 2012-6-7-3 YusunxHuangjin tea No. 2 green with 910 95.5 chestnut 94.0 ess and 95,5 92.5 9370 8 bright relatively bright fragrance mellow neat and even

Table 8 Examination on High-Quality and Strong-Fragrance New Lines for Black Tea

Interior qualit Total

Sequence 9 y Total

Variety Combination name Shape scor number : rank

Tea liquor color Fragrance Taste Infused leaf e

C.sinensis Bit tight, black Strong Red / Bit red Sweet and 94 1 2012-6-21-5 cv.BixiangzaoxWunluza brown, relatively 91.0 93.5 flower 96.0 95.0 bright, neat 9400 1 bright mellow 0 0 moist fragrance and even

Sweet

C.sinensis Bit tight and thin, Relatively . } Bit red fragrance Bit sweet 92 2 2012-6-17-9 cv. BixiangzaoxBaojing bit black, having 83.5 93.5 . 95.0 93.5 red and 93.73 2 bright with flower and mellow 0

Huangjin tea No. 1 hairs bright fragrance

Bit sweetness

Bit tight, having

Fuding White Strong and . gold hairs, Bit red 94 3 2012-6-53-12 TeaxBaojing Huangjin 920 94.0 gardenia 96.0 meilow, 93.0 Red bright 93.70 3 reddish brown bright 0 tea No. 1 fragrance strong with bit moist gardenia fragrance

Bit tight, reddish brown with Red

Fuding White TeaxLong- Bit red Honey Sweet and 94 4 2012-6-58-4 relatively moist, 93.0 92.0 94.5 94.0 bright, neat 93.68 4 leave Baihao bright fragrance meilow 0 gold hair and even appearing / Tight, black Bit sweet,

C.sinensis Strong Red . brown with bith Bit red having . 94 2012-6-32-6 cv.BaihaozaoxZhuyedi 93.0 / 93.0 flower 95.0 93.0 bright,neat 93.60 5 moist, having bright flower .0

No. 9 fragrance and even gold hairs fragrance 21

Relatively tight, Bit strong

TieguanyinxBaojing Bit red Bit sweet 94 6 2012-6-28-5 black brown, 92.0 93.5 flower 94.5 93.5 Red bright 93.43 6

Huangjin tea No. 1 | bright and mellow a relatively moist fragrance

Bit tight, reddish Sweet Relatively : ZhuyeqixBaojing brown with bith orange red, fragrance Strong with red 93 7 2012-6-52-1 92.0 94.0 95.5 92.0 93.23 7

Huangjin tea No. 1 moist, having bright with flower sweet bright,neat .5 gold hairs fragrance and even

Tight, black Red bright,

Bit strong Strong, brown with bith relatively 93 8 Zhuyegi CK 93.0 Red bright 93.0 sweet 93.0 slightly 92.0 9270 8 moist, having neat and .0 fragrance astringent ghold hair even

Note: processed Black Tea and Green Tea 2012-6-28-5 and 2012-6-52-1° both belong to a high-quality and strong-fragrance type, so totally 13 high-quality and strong-fragrance new lines are provided in Table 7 and Table 8. 22

Table 9 Yield Table for High-Yield New Lines

Sum Growth vigour (Year

Compar

Seque of 2019/month/day) ed with ~~ = nce Line yield / Name Fuding numbe (variety) s per /

White 3/6 3/23 6/4 r plot

Tea (+%) (kg)

C.sinensis 1 2012-6-55-4 cv.BaihaozaoxTieguanyi 28.99 112.69 Good Good Excellent- n 2012-6-64- Excelle Excellent 2 ] WuniuzaoxYabukita 29.29 114.89 t Excellent 3 n +

C.sinensis 2012-6-12- Excelle 3 4’ cv.BixiangzaoxTieguanyi 29.24 114.53 t Good Excellent- n n

TieguanyinxBaojing Excelle Excellent Excellent 4 2012-6-28-1 33.38 144.9

Huangjin tea No. 1 nt + +

Fuding White 2012-6-44-7 34.85 155.69 Good Excellent Excellent-

TeaxJinxuan

Fuding White Excelle 6 2012-6-43-2 37.7 176.6 Excellent Excellent

TeaxWuniuzao nt

YusunxHuangjin tea No. 7 2012-6-7-3 ) 32.57 138.96 Good Good Good+

C.sinensis 2012-6-12- Excelle 8 15 cv.BixiangzaoxTieguanyi 38.42 181.88 t Excellent Excellent- n n 2012-6-12- Excelle 9 5 cv.BixiangzaoxTieguanyi 44.15 223.92 t Excellent Excellent n n

FudingxBaojing Huangjin Excelle 2012-6-53-3 27.68 103.08 Excellent Excellent tea No. 1 nt 2012-6-64- Excelle 11 12 WuniuzaoxYabukita 31.28 129.49 t Excellent Excellent- n

Excelle 12 2012-6-66-5 YulvxWuniuzao 31.43 130.59 t Excellent Excellent n

C.sinensis 2012-6-17- Excelle 13 30 cv.BixiangzaoxBaojing 27.75 103.61 t Excellent Good+ n

Huangjin tea No. 1 23

C.sinensis 2012-6-17- 14 0 cv.BixiangzaoxBaojing 27.56 102.21 Good Excellent Good+

Huangjin tea No. 1

Fuding White Excelle 15 2012-6-44-5 28.58 109.65 Excellent Excellent-

TeaxJinxuan nt

C.sinensis Excelle 16 2012-6-39-6 45.56 234.28 Excellent Excellent cv. BaihaozaoxYabukita nt

Excelle Excellent 17 2012-6-66-6 YulvxWuniuzao 334 182.51 t Excellent n +

C.sinensis

Excelle Excellent Excellent 18 2012-5-32-8 cvBaihaozaoxZhuyegi 37.75 212.14 1! n + +

No. 9 2012-6-53- FudingxBaojing Huangjin Excelle Excellent Excellent 19 40.65 174.54 18 tea No. 1 nt + +

HuangqixHubei Tea No. Excelle Excellent 20 2012-6-73-3 36.95 111.3 Excellent 1 nt +

Excelle Excellent 21 2012-6-60-2 Anhui No. 1xYusun 39.5 173.51 t Excellent n +

Yunnan black long

Excelle 22 2012-6-22-2 leavex C.sinensis 38.58 191.75 t Excellent Excellent n cv.Baihaozao

C.sinensis

Excelle Excellent 23 2012-6-67-3 cvBaihaozaoxAnhui No. 43.22 219.22 t Excellent n + 1

C.sinensis

Excelle 24 2012-6-9-13 cv.BixiangzaoxLongjing 33.19 123.73 t Excellent Excellent n

Long-leave

C.sinensis

Excelle 25 2012-6-9-21 cv.BixiangzaoxLongjing 34.86 106.41 t Good Excellent- n

Long-leave

Note: the yields of 25 new lines all exceed that of Fuding White Tea. 10.6 Production test. 18 lines are selected from the above 42 high-quality, strong-fragrance, high-amino-acid and high-yield fine lines for expanding propagation, and a production test and a regional trial are assigned and listed in a registration plan (Table 10).

Various technical features of the above embodiments may be arbitrarily combined. For simplicity of explanation, all possible combinations of various technical features of the above embodiments are not described. However, as long as the combinations of the technical features are not contradictory, it should be considered as a scope described in the description. 24

Table 10 New Lines to Be Registered

Sequence

Line Source Row Ear number 1 2012-6-12-15 C.sinensis cv.BixiangzaoxTieguanyin 241 12050 2 2012-6-12-3 C.sinensis cv.BixiangzaoxTieguanyin 246 12300

C.sinensis cv.BixiangzaoxBaojing 3 2012-6-17-9 365 18250

Huangjin tea No. 1 4 2012-6-21-5 C.sinensis cv. BixiangzaoxWuniuzao 287 14350

Yunnan black long leavex C. sinensis 2012-6-22-2 505 25250 cv.Baihaozao 6 2012-8-28-1 TieguanyinxBaojing Huangjin tea No. 1 198 9900 7 2012-6-28-5 TieguanyinxBaojing Huangjin tea No. 1 30 1500 8 2012-5-32-6 C.sinensis cv.BaihaozaoxZhuyeqi No. 9 483 24150 9 2012-6-44-7 Fuding White TeaxJinxuan 405 20250 2012-6-52-1' ZhuyeqixBaojing Huangjin tea No. 1 141 7050

Fuding White TeaxBaojing Huangjin tea 11 2012-6-53-12 104 5200

No. 1

Fuding White TeaxBaojing Huangjin tea 12 2012-6-53-18 487 23350

No. 1 13 2012-6-55-4 C.sinensis cv.BaihaozaoxTieguanyin 309 15450 14 2012-6-62-1 Baxian TeaxHuangjin tea No. 2 54 2700 2012-6-66-6 YulvxWuniuzao 506 29800 16 2012-6-67-3 C.sinensis cv.BaihaozaoxAnhui No. 1 318 15900 17 6/7/2012-3 YusunxHuangjin tea No. 2 228 11400 18 2012-6-73-3 HuanggixHubei Tea No. 1 449 22450

The above embodiments only express several implementations of the present invention, are described relatively specifically and in detail, but shall not be construed as limitations to the scope of the patent of the present invention. It should be noted that several variations and modifications 5 may also be made without departing from the concept of the present invention for those skilled in the art, and these variations and modifications belong to the protection scope of the present invention. Therefore, the protection scope of the patent of the present invention shall be defined in accordance with the appended claims.

Claims (2)

CONCLUSIONS A method for artificially crossing tea plants, which method comprises the following steps: 1.1 selecting tea plant varieties as hybrid parents based on strong odour, high amino acid content, high resistance and high yield, with a high fruiting rate and/or viability of the relevant pollen; 1.2 intersection of the in step 1.1. selected male and female parent plants that differ in genetic basis and are complementary in one or more traits; 1.3 artificial hybridization pollination 1.3.1.cultivation of the parent plants: the cultivation and management of a female hybrid parent plant should be reinforced in winter in the last year; old branches, weak branches, diseased branches, diseased branches and water shoots are cut off; and more phosphorus and potash fertilizers are applied, so that flowering and fruiting are increased in the following year; collection and storage of pollen: on the day before the artificial hybridization, the budding flowers are picked and placed in a bag; the bag is placed in a dryer and the pollen is prepared the next day by light tapping; and if pollen is brought in from other places or if the flowering stage of the female parents is late, the beaten pollen is kept at a low temperature (4°C or -20°C) by means of a desiccant; method of pollination: monc crossing and pollination with mixed pollen from several male parents are used; artificial pollination: budding flowers in the lower and middle part of the tea plant if female parents in full bloom are selected and subjected to artificial pollination at 7-9 am on a sunny day with no wind or before 11 am on a cloudy day; on the same day, the budding flowers, after being emasculated, are immediately pollinated and packed in a bag; the bag being removed after about a week; 1.4 selection of hybrid varieties 1.4.1.seed harvest: about a week before and after the frost period in the following year, when 70-80% of the husks are dark brown or green-brown and lost their luster, and 4-5% of the tea fruits burst, the seeds are harvested; 1.4.2. sowing and raising seedlings: hybrid seeds are sown over in winter and sown in spring the following year, and the tea fruits should be moistened with fine sand; before sowing and propagation of the hybrid seeds, the hybrid seeds should be soaked for 3-5 days in a solution of 3% carbendazim with a water change once a day, the infertile seeds removed, and the remaining seeds washed with clean water and air-dried for final sowing; sowing uses single seed drilling with a planting distance of 10 cm x 3 cm and a depth of 3 - 5 cm; after sowing, straw with a length of 2 - 3 cm is laid, after which the straw is covered with a mulching film to improve the soil temperature and retain moisture; when a temperature of about 20°C is reached in March-April, the foil is laid open; 1.4.3.hybrid tea seedlings in the F1 generation are selected according to A Comprehensive Evaluation Method For Growth Vigor Of One-Year-Old True Seedlings Of Tea plants by Saijun Li et al; 1.4.4. identification and screening of selective grow rooms; 1.4.5.Identification of the reproductive capacity of seedling cuttings 1.4.6.comparative test of varieties; 1.4.7.expanding propagation for production, experimental planting and regional trials. The method of artificially crossing tea plants according to claim 1, wherein 10 new hybrid varieties are selected and bred, 18 new lines of high quality and high yield are registered, and 77 new lines, 302 hybrid strains in F1 generation and 955 individual plants in F1 generation are tested.