THE BASICS OF CORN BREEDING: INBREDS AND HYBRIDS

Corn, a tropical crop, originated approximately 8,000 years ago in Central America. Many different types of corn evolved with the help of indigenous people who were the first corn breeders. The ultimate goal of corn breeding in Canada is to improve the adaptation of corn to temperate and short season environments. Improved adaptation results in higher yields and better quality. The development of a new corn hybrid is a slow and costly process. New hybrids must possess improved yield, standability, pest resistance and tolerance to various stresses. To achieve this requires the expertise of breeders, entomologists, pathologists, physiologists and many other specialists. There are five major steps in the development of a commercial corn hybrid: 1. 2. 3. 4. 5. identification of appropriate parental germplasm; development of superior inbreds from the parental populations; evaluation of the inbreds in experimental hybrid combinations; identification of a superior hybrid combination(s); and multi-location testing of the pre-commercial hybrid(s).

Once developed, extensive seed production and marketing of the hybrid is required. To understand these processes, a basic knowledge of the structure of the corn plant and general breeding processes is required. Corn has separate male and female flowering parts (Figure 1). The tassel produces pollen and is the male flower; the ear is the female flower. A typical hybrid corn ear consists of several hundred kernels attached to the cob (rachis) and is surrounded by modified leaves that form the husk. Each kernel starts as an ovule and has its own silk that grows out of the husk at the top of the ear. When the tassel is fully emerged from the upper leaf sheath, pollen shed begins from the middle of the central spike of the tassel and then spreads out over the whole tassel. Pollen grains are borne in anthers that open up under appropriate weather conditions. Pollen shed from the tassel usually begins 2-3 days before silk emergence and can continue for several days afterward. Pollen grains are very light and can be carried considerable distances by the wind. Pollen shed will stop when the tassel is too wet or too dry. Pollen grains are only viable for 18-24 hours. The silks are covered with fine, sticky hairs that catch and anchor the pollen grains. Within minutes after landing on the silks, the pollen grain germinates and a pollen tube grows down the silk to fertilize the ovule or potential kernel. This usually takes 12-28 hours. Under good conditions, all silks will emerge and be ready for pollination within 3-5 days. There is usually more than enough pollen

since a given tassel can produce up to 5 million pollen grains (roughly 5,000 per silk), so problems generally occur when there is poor synchronization between silk emergence and pollen shed. The greatest impact on grain yield occurs when environmental conditions are unfavourable during pollination time. Corn, because it has separate male and female flowering parts, is a naturally cross-pollinated plant. Kernels can be pollinated by pollen from neighbouring plants; therefore, care must be taken in a breeding program to ensure that pollen from the appropriate tassel fertilizes ovules on the appropriate ear. This is often achieved by conducting hand-pollinations. As soon as ear shoots are visible in the leaf axils of a plant, a small paper 'shoot-bag' is placed over the ear shoot (Figure 2); this allows the ear to continue growing and the silks to emerge but prevents any pollen from falling on the silks. When the tassels are shedding pollen, a paper bag is placed over the tassel and stapled or paper-clipped at the base of the tassel to trap any pollen that is shed. The next day the tassel bag is removed and quickly placed over a covered ear that has silks, taking care to quickly remove the shoot-bag on the ear. Alternatively, a see-through glassine bag can be placed on the tassel and shaken to allow pollen to collect in one corner of the bag; this corner is then ripped off and the pollen is deposited on the silk (Figures 3a, 3b) and then the whole ear is rapidly covered with a brown paper tassel bag. With both techniques, the tassel bag is pulled around the stalk, stapled and shaken to assist the pollen grains in falling on the silks (Figure 3c). A plant is 'self-fertilized' (also referred to as 'selfing' or inbreeding) when the pollen from a tassel is placed on the silks of the ear of the same plant (Figure 4). A plant is 'cross-fertilized' or 'crossed' when the pollen from a tassel is placed on the silks of a different plant (Figure 4). Of the millions of hand pollinations made by corn breeders, only an extremely small amount ever result in a superior inbred that will be used in a commercial hybrid. From 1850-1910, North American corn breeders developed higher yielding openpollinated corn varieties. The plants in a given field were allowed to shed pollen and no silks were covered, resulting in a mix of cross and self-pollinated kernels on each ear. The best plants would be selected and their ears (usually the largest ones in the field) would be kept to use as seed the next year. The resulting plants were gradually improved for agronomic traits, but were variable in plant height, ear height, maturity, etc.due to the continuous cross-pollinations. In the 1920's, the concept of hybrid vigour (heterosis) was discovered. If corn plants were continually self-pollinated for six or more generations, the plants became smaller (due to a decrease in vigour) but more uniform for all traits. Selection can be made every generation for specific traits such as pest resistance, plant or ear type, ear size, etc. This produces what is called an inbred line or inbred. The development of an inbred can be accelerated with the use of winter nurseries in warmer southern climates. An inbred is genetically uniform for all traits and will breed true to form. Hybrid vigour is demonstrated two inbred lines from unrelated backgrounds are cross-

pollinated (Figure 5). The offspring of such a cross will have a larger ear and will be a more robust plant. It is also uniform for most traits. There are many theories to explain the phenomenon of hybrid vigour, but few are adequate to explain everything. If an ear of hybrid corn is self-pollinated, the resulting progeny will be not be genetically uniform, will be variable in height and have lower yields. This is why farmers must buy their hybrid corn seed each year and should not keep the seed from a field of hybrid corn to plant next year. Development of inbreds takes about 75% of the breeding resources in a corn breeding program. The largest effort is spent evaluating whether or not the inbred when crossed to another inbred which is called a 'tester', will produce a hybrid with high yield and good agronomic characteristics. This is called evaluating the combining ability of the inbred. The cross or hybrid is called a 'testcross'. The field performance of this testcross is extensively evaluated in replicated multilocation field trials. Inbreds with superior testcross performance are retained and advanced to the next generation. It would be great if selection could be done at the inbred level since expenditures for testing could be reduced considerably if the performance of the inbred on its own could predict the performance of the hybrid testcross. This happens for some traits such as earliness, plant height and disease resistance but unfortunately not for yield. Besides having good combining ability, an inbred line must be easy to maintain and to cross if hybrid seed production costs are to be kept low, and a consistent production of high yields of uniformly good quality seed is to be maintained. Pure stocks or breeder's seed of inbred lines used in commercial hybrids must be maintained by hand-pollination. For production of hybrids, inbred seed can be increased in isolated fields which must be planted a minimum of 200 m from any other corn crop. For each commercial hybrid, large quantities of seed of its parental inbred lines and the cross must be produced to ensure an adequate supply of seed is available for growers. Hybrid seed is produced by planting the 'female' and 'male' inbred lines together in a field. The choice of which inbred to designate female and which to designate male depends on the ear and tassel characteristics of each, usually the female has higher seed yield and the male has good pollen production. The ratio of female to male rows varies with different seed companies. Differential planting dates can be used to ensure synchronization between male and female flowering. Female rows are mechanically and/or hand-detasseled shortly after the tassel has emerged from the uppermost leaf sheath and before any female plants begin to shed pollen. At harvest, commercial seed corn fields are usually harvested by a picker/husker and the husked ears are sorted to remove off-type ears and trash. The seed is dried, shelled, cleaned and graded. Germination tests are also carried out and the seed is treated with a fungicide before packaging. Today, more than 80% of corn grown in North America is a single-cross hybrid of this nature. The remaining 20% consists of double, three-way and modified

(related-line parents) crosses. Corn yields have increased in North America from approximately 1.3 Mg/ha in 1930 to 8.7 Mg/ha in 1994 or approximately 0.078-0.110 Mg/ha per year. This steady increase is due to improved hybrids, increased use of fertilizers, better weed control, higher plant densities and better management.

Fig. 1 (click the image to see a larger view). The basic structure of a corn plant. The separate male (tassel) and female (ear) flowering structures make corn a naturally cross-pollinated plant.

Fig. 2 (click the image to see a larger view). The ears on these corn plants have been covered prior to silking with a white glassine 'shoot-bag'. This bag prevents any pollen from fertilizing the silks.

Fig.3a (click the image to see a larger view). The corner of a glassine tassel bag containing pollen is ripped off

Fig.3b (click the image to see a larger view). The pollen is quickly dropped onto the silks (the larger particles are anthers from the tassel).

Fig.3c (click the image to see a larger view). After pollination, a brown paper bag is placed over the ear and the ends of the bag are stapled around the stalk. Fig. 3. Pollinating corn. The corner of a glassine tassel bag containing pollen is ripped off (a) and the pollen is quickly dropped onto the silks (b) (the larger particles are anthers from the tassel). After pollination, a brown paper bag is placed over the ear and the ends of the bag are stapled around the stalk (c)

Fig. 4 (click the image to see a larger view). Self pollination vs. cross pollination. A plant is selfed when it is fertilized with pollen from its own tassel; a plant is crossed when it is fertilized with pollen from another plant's tassel.

Fig. 5 (click the image to see a larger view). A hybrid is produced when two compatible inbreds, usually of different genetic backgrounds, are cross-

pollinated. The hybrid plant is much larger and more robust than either of its inbred parents. This is referred to as hybrid vigour or heterosis.

How Rice is Bred

The rice breeding team uses hundreds of different rice lines from all over the world to produce the varieties which are grown by Australian farmers. These lines are painstakingly crossed using a technique called approach crossing and are raised in small numbers in the glasshouse. They are carefully monitored for their ability to produce the characteristics the breeders are selecting for. The relatively small amount of seed gathered from these glasshouse-reared rice plants is used to sow trial plots where the many different crossbreeds are again monitored for vigour, disease and insect resistance, water use, growth habit and grain quality, under field conditions. It takes breeders on average 10 years to develop a new rice variety. Development begins with an annual meeting between NSW Agriculture rice breeders and rice marketing experts to discuss what grain characteristics need to be developed to meet the demands of new markets both in Australia and overseas. These decisions are usually difficult because the breeders and marketing experts are trying to predict consumer preferences for the combinations of grain and cooking quality, many years into the future. Once the desired make up of the new rice is determined, breeders screen the different genotypes, or strains of rice, held in the Rice Breeding Program collection to find plants that might match the market requirements or are suitable for use as parent plants. The program holds many thousands of genotypes which are drawn from all over the world and stored as seed. Seed from the selected genotypes are grown in the glasshouse in preparation for approach crossing which is the first phase of the plant breeding operation. Related Links: Grain Quality, Approach Crossing, Plant Breeding Operation

A range of factors combine to affect the quality of rice from the paddy stage through to the quality of the finished product. In general terms, these factors are: the genetic make-up of a particular variety environmental factors, e.g. weather and farming practices the effects of grain handling, (harvesting, storage, drying, processing and distribution).

The quality of rice is generally a reflection of the desired use of the rice product. Uses can vary from an accompaniment for Asian foods, to specialised rice recipes such as risotto or sushi, to an ingredient for breakfast and snack foods. Standard quality tests are also applied to paddy rice, as the grain is called before the hull is removed. The quality tests applied to paddy rice when it is delivered for storage relate to varietal or grain type purity, moisture content and contamination by physical, chemical and biological materials. Additional grain quality criteria are applied during the various stages of milling and processing. These include: milling yield from paddy, brown or whole grain rice percentage of broken grains degree of polishing. Rice grain is composed of starch, protein and lipids (oils) and the amounts of each in the grain underlie the physical attributes of rice which are: . anatomy grain length and width grain weight hull & bran colour whiteness chalk cracking % whole grain and cooking and eating characteristics of rice. These are: Amylose content Gelatinisation temperature Viscosity Texture Flavour and aroma The Grain

Grain Composition and Physical Attributes Images Rice grain crosssection | Panicle Position | Paddy Rice

Anatomy

Grain Quality Factors

Grain Length, Width and Weight

Whiteness

Cracking

Chalk

Hull & Bran Colour

% Whole Grain

Nutrition

Classifying Rice

Wet-Dry

Physical Attributes

Cooking and Eating Characteristics

Amylose Content

Gelatinisation Temperature

Viscosity

Texture

Flavour and Aroma

Comparison of Cooking Characteristics Table

Approach Crossing
The plant breeder modifies one of the chosen parent plants at the start of the approach crossing process by delicately snipping away part of its tiny flowers, known as the florets. Only the female is altered in this way and the plants are partially covered by plastic and placed together in the glasshouse. Approach crossing ensures that pollen from the male plant fertilises only the female it is being crossed with. Plant Breeding - Approach Crossing

Step 1 Suitable parent plants are selected from the thousands of varieties held by the NSW Agriculture Rice Breeding Program

Step 2 The florets of the rice plant chosen to be the female are cut to remove the anthers, the part of the plant where the pollen is stored.

Step 3 The panicles, or flowering stems, are drawn together inside a plastic bag to help make sure the pollen falls from the male to the female parent.

Step 4 Seed is ready for harvest one month later

Detailed Operations of the Rice Breeding Program

Plant Breeding - Detailed Operations of the Rice Breeding Program Year Approach crossings are made in the 1 glasshouse in October and March Generation Year Plant numbers are increased in the 1 2 glasshouse, usually during summer season Bulk populations are sown in field, often at two locations. Generation Year Single flower bearing stems, or panicles, are 2 3 selected. Grain is visually screened for shape, chalk and breakage 1 million plants 150-300 crosses

Seed from single panicles are sown as a panicle (short) row and records of heading date are kept. Generation Year Selected single panicles from the population 3 4 of the rows are bulked together in a single packet (no record is kept of the source row). Each selected panicle is screened for grain shape, chalk and breakage. Single rows of seed from selected panicles are sown and a record of flowering date is made. Selected rows are harvested in bulk and Generation Year screened in the laboratory for grain shape, 4 5 chalk and breakage. Selected rows are also screened for amylose and early indications of cooking time. A single unreplicated plot is sown. Establishment count, heading date, height and hairiness (or pubescence) are recorded. All rows are harvested and yield recorded. Rows are tested for moisture content and quality samples are taken and dried. First cull on field performance occurs. Generation Year Preliminary quality evaluation is conducted 5 6 on brown rice Milling quality test is performed White rice tested for chalk, colour, shape and size Selected rows tested for cooking and eating texture Panicles are usually selected from the plots to commence seed multiplication Selected rows are sown in replicated plots Generation Year four replicates at Yanco and two replicates 6 7 at Deniliquin. 400 varieties 4,000 plots

35,000 rows

Best lines are sown in preliminary district trials. Generation Year 7 8 Seed multiplication takes place at Yanco Quality evaluation of selected lines is more intensive (Serial samples for milling quality and appearance, cooking and eating texture). Second District Trials Generation Year Intensive quality evaluation continues 8 9 Seed multiplication takes place at Jerilderie (producing 0.5 tonnes) Usually a second year of district evaluation Generation Year 9 10 Seed multiplication takes place at Jerilderie (producing 25 - 50 tonne) Generation Year Commercial production of 150ha (may be 10 11 released in this season). 1 variety 40 varieties