lnvention and Utilization of Glutinous Sorghum Male Sterile Line 45A with High Starch and High Combining Ability

Guoxiang DlNG，Ganlin ZHAO，Guomin CHEN，Xianlin Nl，Tianpeng LlURice and Sorghum Institute，Sichuan Academy of Agricultural Sciences，Luzhou 646000，China

lnvention and Utilization of Glutinous Sorghum Male Sterile Line 45A with High Starch and High Combining Ability

Guoxiang DlNG*，Ganlin ZHAO，Guomin CHEN，Xianlin Nl，Tianpeng LlU
Rice and Sorghum Institute，Sichuan Academy of Agricultural Sciences，Luzhou 646000，China

45A is a glutinous sorghum male sterile line with high starch and high combining ability bred by Rice and Sorghum Institute of Sichuan Academy of Agricultural Sciences in 1998，it is a coeno-species taking non-glutinous maintainer line TL169-239B which bred by Tieling Institute of Agricultural Sciences in 1991 as the female parent and glutinous maintainer line72B bred by the authors'institute as the male parent，then，based on the backcross breeding between glutinous single plant chosen from F2segregation population and Tx623A.There are ten hybrid sorghum varieties which already have been examined and approved by national and above provincial（municipal）level units；the patent of this breeding method has been authorized （the patent number:ZL 2012 1 0129155.6）；45A is protected by the Right of New Varieties of Plants， MOA， P.R.China （the variety right number: CNA20090576.1）.In this paper，the breeding process of 45A and the characteristics of 45A sterile line and its hybrid sorghum were summed up，what's more，the technical key points of high-yielding breed of 45A and the production of hybrid sorghum seeds were introduced，to provide data for further popularizing the sterile line.

Sorghum；Male sterile line；Starch；Combining ability；Glutinous；Invention；Utilization

W ith the large extension of non-glutinous hybrid sorghum in sorghum production，there were such problems as poor quality of liquor-making and low yield of common glutinous sorghum varieties，therefore，the utilization of hybrid with good quality and high yield is the only way for liquor-making sorghum industry development，in which the breeding of glutinous sorghum parental lines with high starch and high combining ability is the key to success.For this，based on studying the hybrid vigour of glutinous and non？glutinous sorghum［1］，and by hybridizing glutinous and non-glutinous sorghum from different distant geographic environment，the authors aimed to breed glutinous sorghum male sterile line with high starch and high combining ability，and overcome the technical difficulty of unifying good quality and high yield，as well as invent new liquor-making sorghum variety with the good characteristics of good quality，high yield，strong resistance and extensive adaptability，thereby，ensuring the food security and sustainable development of liquor-making industry in China.

Breeding Process

In1991， some batches ofsorghum materials were introduced from Tieling Institute of Agricultural Sciences and then planted in Luzhou，TL169-239B showed the features of large grain，long spike，red grain，nonglutinous and moderate loose spike，etc.，but had the defects of higher plant and longer growth period.Glutinous sorghum maintainer line 72B bred by Rice and Sorghum Institute of Sichuan Academy of Agricultural Sciences had the features of glutinous endosperm，smaller 1 000-kernel weight，short spike，white grain and light grain weight per spike；and its parents had better supplementary，the genetic basis was extensive，F1generation showed the characteristics of larger 1 000-kernel weight and heavy grain weight per spike，which conformed to the breeding objective of long-spike hybrid sorghum maintainer line.Since F2generation which hybridized from TL169-239B/72B，iodine and potassium iodide chromogenic reaction had been adopted to detect glutinous grain；for F3generation，single plant with excellent agronomic trait and high disease resistance was chosen to cross with Tx623A；for F4，F5and F6，based on the identification of resistance and quality，the restorable ability and combining ability of selected single plant or strain were measured，then according to the advantages of measured combinations，the strains with strong combining ability and red grain were chosen to breed and measure，which was named as 45A，and it passed technical appraisal in 2002.

Table 1 45A breeding flow chart

The Characteristics of 45A Sterile Line and Its Hybrid Sorghum

The stability of hereditary feature and agronomic trait of 45A

The agronomic trait and hereditary feature of 45A are homozygous and stable.By identifying，there is no difference among individuals，and the agronomic trait is the same.

Morphological characteristics Coleoptile and seedling are green，vein is waxy，the number of blades is about 20-21，the plant height is 125-135 cm，the spike is moderate loose type and long spindle shape，spike length is 32-38 cm，grain weight per spike is 50-55 g，spike has soft brown shell and red grain，and the 1 000-kernel weight is about 24 g.

Biological characteristics In Luzhou，the initial sowing time of 45A is in April and the growth period is 120-125 d，there is generally 90-95 d from sprouting to half bloom.The sterility is stable，the anther is white and wizened，thus the fertility rate is 100%；the stigma of pistil is feathered and milk white with high exsertion rate，the affinity is high and the flowering phase is long，allogamous time is 7-9 d，lack of sun during the meiophase can lead to abortion phenomenon of parts of flowers.The physiological races with Sphacelotheca reiliana（Kühn.）Clint. are vaccinated against leaf spot disease.

Quality 45A grain contains 8.23% crude protein，75.56% total starch，74.13%amylopectin which accounts for 98.12% of total starch，3.22% crude fat，0.41%tannin，and glutinous endosperm.

High combining ability of 45A

In general，45A has high combining ability，high yield and fine quality，thus its hybrid sorghum is the organic bond of high yield，fine quality and wide adaptability.

Table 2 showed that 45A had high combining ability，the general combining ability of grain weight per spike and 1 000-kernel weight was remarkably high.45A glutinous endosperm had fine quality，the combining ability of total starch and amylopectin was high，which had become the backbone parents of three-line hybrid sorghum combination，and it formed a series of fine hybrid sorghum combinations with good quality，high yield，good yield stability，wide adaptability and strong stress resistance.Up to now，therehave been ten 45A series combinations authorized by national and above provincial（municipal）level units（see Table 2），including 6 combinations authorized by national units and 4 combinations authorized by provincial（municipal）level units，in which 4 combinations were recommended as main varieties by each agricultural sector.

Table 2 Authorized hybrid sorghum combinations of 45A and others

Good restorability of 45A Through crossing with restoring lines35R，07221，3560，Jinliang 5，Luhui 1，LZ2R and 5-26，the setting percentage of hybrid F1was above 88%，and the average rate was 91.3%，which was obviously higher than existing glutinous sterile line（Table 3）.

High combining ability of 45A yield

The effect value of general combining ability of the parents reflects the average performance of a character in a series of hybrid combinations.If the value is high，indicating that beneficial minor genes for the character are more，additive gene effect is greater， and the genetic character is stable.Ni Xianlin et al.adopted incomplete diallel cross test in 2011［2］（Table 4），the analysis showed that the general combining abilities of 45A and grain weight per spike，1 000-kernel weight，spike length were higher，the relative effect values of general combining ability were 2.43，1.24 and 1.52，respectively，showing that varieties matched with 45A had high yield per plant and heavy 1 000-kernel weight as well as long ear length.

High general combining ability of total starch and amylopectin contents in 45A Ding Guoxiang et al. used incomplete diallel cross test to analyze starch content and combining ability of its components in 2011［3］（seen in Table 5），proving that effect values of general combining ability of total starch and amylopectin contents in sterile line 45A were high，namely，the relative effect values were 1.238 and 2.915，respectively，which was consistent with Zhao Jing’s research results［4］（Table 6）.The research showed that 45A was a good parent for total starch and amylopectin improvement.

High yield and wide adaptability of hybrid sorghum crossed with 45A

There were ten hybrid sorghum varieties crossed with 45A examined and approved in Sichuan，Shanxi and Chongqing，yield of Lunuo No.3［5］，Lunuo No.8［6］，Lunuo No.9［7］and Lunuo No.10［8］regional trial reached more than 5850 kg per unit area，which increased by above 35%than conventional sorghum variety—Qingkeyang sorghum.Lunuo No.1 yield was 6 840.0 kg/667m2and increased by 25.6% comparing with two-line glutinous hybrid—Liangnuo No.1.Yield of Jinza 23［9］，Jinza 104 and Jinzhong 0922 regional trial was above 8 000 kg/667 m2and increased by 12.1%-15.1%comparing with Jinza 12，in which yield of Jinza 23 was 9 832.5 kg/667 m2.In re-gional trials，yield of 6 combinations ranked the first，and the others ranked the 2ndor the 3rd，yielding ability was prominent，thus they were suitable to be popularized in North China and Southern Areas（Table 7）.By the vertical comparison，yield of Lunuo No.1（72A/35R）which authorized in 1993 was 4 405.5 kg/667 m2，however，yield of glutinous sorghum crossed with 45A increased 1 500 kg/667 m2than Lunuo No.1.

Table 3 The percentage of bagging selfing of hybrid combinations between 45A and part of restoring lines　%

Since the breeding of 45A female parent hybrid sorghum，it already has been popularized all over China due to its high yield，good quality and strong disease resistance.In 2008，the yield of Lunuo No.8 was measured in Jiangyang District of Luzhou City，yield of the first cropping sorghum was 8 944.5 kg/667m2，ratooning sorghum yield was 6 270.0 kg/667 m2，thus total yield was 15 214.5 kg/667 m2.In 2009，the yield of Lunuo No.8 was measured in Zhuji of Zhejiang and the first cropping yield was 9 672.8 kg/667 m2.In 2010，experts inspected Jinza 23 on site，the yield was 15 439.5 kg/667 m2，which set the new domestic record of sorghum per unit yield.

Strong ratooning ability of hybrid sorghum crossed with 45A，good yielding ability of ratooning sorghum As seen in Table 8，the experiment was carried out in Jiang’an Agro-technical Station in 2008，it showed that Lunuo No.3，Lunuo No.8，Lunuo No.9 and Lunuo No.10 had strong regenerative capacity，and grew quickly and well，ratooning sorghum yield reached more than 5 400 kg/667 m2，the growth period was short，its yield-increasing effect was prominent than conventional sorghum.In Zhuji of Zhejiang，yield of Lunuo No.8 exceeded 12 000.0 kg/667 m2in the two harvests for five years，thus achieving one seeding and two harvests.

Table 4 The relative effect values［3］of general combining ability of 45A and other parental lines

Table 5 The relative effect values［3］of general combining ability of starch contents in 45A and other parental lines

Table 6 The relative effect values［4］of general combining ability of liquor-making quality of 45A and other parental lines

High starch content and strong glutinous feature of 45A，and good liquor-making quality of its hybrid sorghum combinations

45A is a glutinous sterile line with the characteristics of good quality and large grain.Some Maotai-flavor liquormaking enterprises believed that conventional glutinous red sorghum with small grain had high quality.From Table 9，45A and its corresponding maintainer line 45B had particularly outstanding integrated quality.Andmeasured by Luzhou Institute of Brewing Technology，it found that 45B had oval grain and reddish brown appearance，the 1 000-kernel weight was 24.1 g （Luzhou），total starch content was 75.56%，amylopectin content was 74.13%（accounted for 98.12%of total starch），contents of protein，tannin and fat were 8.23%，0.41%and 3.22%，respectively.Therefore，compared with other domestic glutinous maintainer lines，45B achieved extremely remarkable genetic improvement not only in exterior quality character，but also in grain quality character.45B had higher 1 000-kernel weight，effectively coordinating the contradiction between grain weight and quality.

Hybrid sorghum crossed with 45A had high liquor rate and good liquor quality.As shown in Table 10，total starch content of ten hybrid sorghum varieties crossed with 45A was above 73.09% ，and the highest one was 78.69%.Amylopectin content of glutinous combinations occupied 96.20%-97.99%of total starch，and that of glutinous-non-glutinous combinations occupied about 85%of total starch.As seen in Table 11，from Luzhou Institute of Brewing Technology，we knew that the liquor rates of Luzhou-flavor，Fen-flavor and Maotai-flavour brewed from Lunuo No.8 had 1.2-2.0 percentage points higher than conventional glutinous sorghum.Institute of Characteristic Crops of Chongqing Academy of Agricultural Sciences carried out Xiaoqu wine brewing experiment using Lunuo No.8 three times in Yongchuan Wuyi Winery，the results indicated that the liquor rate was 59.5%（57 V/V），which was 8 percentage points higher than northern non-glutinous sorghum，it was a full-bodied wine with pure aroma，tasted mellow and refreshing.Lu Zhou Lao Jiao Co.，Ltd.brewed Luzhou-flavor liquor using hybrid glutinous sorghum and its matching technology，comparing with local glutinous sorghum，its liquor rate improved 1.27%，ethyl hexanoate increased by 12.93%，conversely，ethyl lactate reduced by 22.15%，and comparing with northern non-glutinous sorghum，liquor quality improved significantly.Sichuan Tianzhijiaozi Industrial Limited Company brewed Maotai-flavour liquor using hybrid glutinous sorghum and its matching manufacturing technique，the liquor rate improved 6.26%，and ratio of quality goods improved 11.28%.

The disease-resistance of 45A and its hybrid sorghum

The immunity of 45A against Sphacelotheca reiliana（Kühn.）Clint. Microspecies with Sichuan S.reiliana were different from No.3 microspecies. The natural incidence of S.reiliana of 45A was 0，based on the identification of the inoculation of Luzhou species，in the tested 15 sorghum sterile lines，the vaccinal disease incidence of 13 sterile lines of 45A，3246A，407A，401A，3268A，V4A，AGR1A，0122A，187A，3401A，72A and Ji 2055A was 0，for the immune level of tested bacteria，the vaccinal disease incidence of 18A was 15.19%，and reached moderate resistance level，the vaccinal disease incidence of 0967A was 52.38%，and it had high sensitivity［10］.

Table 7 Yield performance of hybrid sorghum crossed with 45A

Table 8 Comparison on ratooning sorghum yield among different 45A hybrid sorghum varieties

The immunity of hybrid sorghum crossed with 45A against physiological race with Sphacelotheca reiliana（Kühn.）Clint.The vaccinal disease in-cidence of the combinations of 45A with 1045R，1050R，1047R，Hui 1，1181R，0953R，3560R was 0，and reached immune level.In 2008，Longmatan District of Luzhou City introduced the varieties and carried out experiments，the disease incidence of Lunuo No.8，Lunuo No.9 and Lunuo No.10 was 0，that of Qingkeyang sorghum and Liangnuo No.1 were 3.5%and 45.5%，respectively.

Leaf disease resistance of hybrid sorghum crossed with 45A against Mampava bipunctella Ragonot.Based on field investigation，the whole plant leaf of the combinations of 45A with Lunuo No.3，Lunuo No.8，Lunuo No.9，Lunuo No.10 was green when maturing，and they can resist leaf spot disease.Generally，the combinations crossed with 45A belonged to loose panicle type and were little influenced by M.Bipunctella.

Breeding and Seed Production Techniques of 45A

The plant height of 45A is about 130 cm，and the shape of plant leaf is compact，it has erect blade，small angle and thick green leaf color，the number of stem leaf is 20-21，the duration from seeding to heading in Luzhou is 95-105 d，the infertility is stable，the degree of infertility and the sterile rate are both 100%，the stigma is white，and it has strong viability；the panicle type is moderate loose and the panicle length is 32-38 cm，the grain weight per spike is 50-55 g，the grain is oval with bronzing color；1 000-kernel weight is 23-25 g.

The optimum flowering temperature of 45A is 25-30℃，and the humidity is above 80%.During the breeding and seed production process，based on the characteristics of higher stigma exsertion rate，longer stigma vigour，longer flowering period and decentralized flowering period，the yield of 45A breeding seed is generally 2 250-3 450 kg/667 m2 by using cultivation techniques and enhancing field management.In 2012，the average yield of 45A in high-yield breeding demonstration plot of Yuanjiang in Yunnan was 3 000 kg/667 m2，the yield of hybrid seed production was 3 900 kg/667 m2.

Table 9 Comparison on grain qualities of 45A and some sterile lines

Table 10 Comparison on grain liquor-making quality of 45A hybrid sorghum

Key points of 45A high-yielding breeding techniques

The optimum seeding time for 45A breeding in Luzhou is in early April.The seeding time of male parent is 3-5 d later than female parent in the first phase，the difference of leaves is 1.0；in the second phase，male parent is 6-8 d later than female parent，for the optimum flower synchronization，female parent is 1-2 d earlier than male parent.The seedbed should be prepared fully as follows:the seed yield of female parent and male parent are 7.5 and 3.75 kg/667 m2，respectively；enough base fertilizer should be applied in seedbed，mulching film and lagging jack should be used for seedling transplantation with sparse and uniform seeding，and the management should be enhanced to breed strong seedling；the row ratio of male parent to female parent is 1:3-5；in the spacing of the first phase and the second phase，two-nest transplanting is carried out，line spacing and nest spacing of female parent are 60 and 30-33cm，respectively；the spacing between parents is 55-60cm，nest spacing of male parent is 30-33cm；two-plant transplanting per nest is implemented for parents；a sufficient supply of base fertilizer must be ensured，topdressing should be carried out early，phosphorus potassium fertilizers should be increased；moreover，we must pay attention to preventing aphid and M.Bipunctella，and weeding before and during flowering timely andeffectively；after pollinating，male parent should be cut off timely.Finally，threshing must be harvested in time to prevent mix.

Table 11 The experiment result of liquor brewing using Lunuo No.8

Key points of 45A hybrid sorghum seed production technique

To arrange seeding interval between parents properly and ensure the optimum flower synchronization. Based on the characteristics of long florescence and long stigma viability of 45A，female parent should be 1-2 d earlier than male parent.

To breed strong seedling and timely transplant，and realize rational close planting.The seedbed should be prepared fully as follows:the seed yield of female parent and male parent are 9.0 and 3.0 kg/667 m2，respectively；enough base fertilizer should be applied in seedbed，mulching film and lagging jack should be used for seedling transplantation with sparse and uniform seeding，and the management should be enhanced to breed strong seedling；the row ratio of male parent to female parent is 1:5-6；in the spacing of the first phase and the second phase，nest transplanting is carried out，line-nest spacing of female parent is 60 cm×30 cm；the spacing between parents is 55-60 cm，nest spacing of male parent is 30 cm；twoplant transplanting per nest is implemented for parents；a sufficient supply of base fertilizer must be ensured，topdressing should be carried out early，phosphorus potassium fertilizers should be increased；moreover，we must pay attention to preventing aphid and M.Bipunctella.

洗衣这件事，是从小就有的爱好。一年四季无论寒暑，就爱洗衣服，差不多七八岁的时候，便热爱了。逐渐到后来，与其说是热爱洗衣，不如说更爱长久地待在江上，顾漫无言地——棒槌声声，沉浸于寒冷中浑然不觉，这也是消耗生命的一种方式吧，不然，那么多的空闲，如何打发呢？洗完一铁桶衣服，拎着它慢慢走在铺满青石条的巷子里回家，浑身发热——总是利用不停歇的体力活，试图去填满生命里出现的大片空洞，仿佛热血犹在，漂泊而失根的小小生命，一步一步有了方向。

To weed before and during flowering timely and effectively.Firstly，we should forecast flowering phase，ensuring flower synchronization.In flowering phase regulation，we should focus on promotion and take the combination of promotion and control as the supplement.In the stage of growth and development，abnormal，heterochromatic and inter-temporal plants must be removed as soon as possible.

Artificial supplementary pollination.In the forenoon of a fine day，after the drying of dew，pollination will be carried out.

To harvest threshing timely. When the color of 90%grain of spike becomes red and hard，indicating that it can be harvested；after airing 3-4 d，it can be threshed.

To prevent mechanical mixing.In the links of harvesting and dry in the sun，threshing and transportation，we must keep dam，tool and bag clean.

References

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［5］DING GX（丁国祥），ZHAO GL（赵甘霖），LI TY（李团银），et al.Breeding and application of liquor-making sorghum variety Lulo No.10 with good quality and high yield （高产糯质酿酒高粱杂交种泸糯3号的选育与应用）［J］.Seed（种子），2005（10）:76.

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Responsible editor:Nanling WANG

Responsible proofreader:Xiaoyan WU

高淀粉高配合力糯质高粱雄性不育系45A创制与应用

丁国祥*，赵甘霖，陈国民，倪先林，刘天朋
（四川省农业科学院水稻高粱研究所，四川泸州646000）

45A是四川省农业科学院水稻高粱研究所1991年以辽宁铁岭市农科院育成的粳质保持系TL169-239B为母本与本所育成的糯质保持系72B为父本杂交，从F2分离群体选择糯质单株与Tx623A回交转育，于1998年育成的高淀粉高配合力糯质高粱不育系，已育成审定10个杂交高粱品种通过国家和省市审定，其育种方法获国家发明专利授权（专利号：ZL 2012 1 0129155.6），45A获得农业部植物新品种权保护 （品种权号：CNA20090576.1）。该文总结了45A的选育过程及45A不育系和其所配杂交高粱的特征特性，最后介绍了45A高产繁殖和杂交高粱制种技术要点，为该不育系进一步推广应用提供资料。

高粱；雄性不育系；淀粉；配合力；糯质；创制；应用

“八五”国家科技攻关项目“高粱亲本系创造和选育技术研究”（85-002-03）；国家十二五科技支撑计划项目：高淀粉酿造高粱新品种选育及产业化示范 （2006BAD02B03）；国家现代农业产业技术体系：高粱（CARS-06-01-06）。四川省“十五”（2001-12-03-04）、"十一五"（2006-02-02-05）、“十二五”（2011NZ0098-6）攻关项目：酿酒高粱新品种选育。

丁国祥（1962-），男，四川合江县人，副研究员，主要从事高粱育种及栽培技术研究，E-mail：dgx6132@163.com。*通讯作者。

2014-12-20

2015-03-07

Supported by National Science-Technology Project of"the Eighth Five-Year Plan": Study on the Invention of Sorghum Parental Lines and Its Breeding Technology（85-002-03）；National Science-Technology Support Plan Project of the 12thFive-Year Plan: Breeding of Brewing Sorghum New Variety with High Starch and Its Industrialization Demonstration （2006BAD02B03）；National Modern Agricultural Industrial Technical System:Sorghum（CARS-06-01-06）；Sichuan Scientific and Technological Project of"the Tenth Five-Year Plan"（2001-12-03-04），"the Eleventh Five-Year Plan"（2006-02-02-05）and"the Twelfth Five-Year Plan"（2011NZ0098-6）:Breeding of Brewing Sorghum New Variety.

.E-mail:dgx6132@163.com

December 20，2014Accepted:March 3，2015